WO2017173089A1

WO2017173089A1 - Systems and methods for enhancing delivery of diagnostic and/or therapeutic compositions in vivo using electric pulses

Info

Publication number: WO2017173089A1
Application number: PCT/US2017/025035
Authority: WO
Inventors: Govindarajan Srimathveeravalli; Thomas Reiner; Stephen Solomon
Original assignee: Memorial Sloan Kettering Cancer Center
Priority date: 2016-03-31
Filing date: 2017-03-30
Publication date: 2017-10-05

Abstract

Described herein are systems and methods for manipulation of tumors with pulsed electric fields. The systems and methods described herein can be used for theranostic applications for oncology diagnosis and treatment, especially when combined with liposome-delivered drugs.

Description

SYSTEMS AND METHODS FOR ENHANCING DELIVERY OF DIAGNOSTIC AND/OR THERAPEUTIC COMPOSITIONS IN VIVO USI NG ELECTRIC PULSES

Cross Reference to Related Applications

This application claims priority to and the benefit of, and incorporates herein by reference, U.S. provisional Application No. 62/316,454, filed March 31, 2016.

Field of the Invention

[0001] This invention relates generally to enhanced drug delivery to tumor tissue. In particular embodiments, the invention relates to combination of liposomes with electric pulses for drug delivery to dense tumors (e.g., pancreatic, colorectal, breast cancer). In other embodiments, the invention relates to tissue ablation using electric pulses.

Government Funding

[0002] This invention was made with government support under EB016673 awarded by

National Institutes of Health. The government has certain rights in this invention.

Background

[0003] Solid tumors, especially those from ovarian, colorectal, pancreatic cancers, certain forms of breast cancers, sub-types of sarcoma, and melanoma are characterized by high interstitial fluid pressure (IFP). High IFP negatively impacts the penetration and distribution of therapeutic agents within these tumors, leading to poor or low bioavailability of these agents. Unsurpri singly, high IFP in a tumor is correlated strongly with poor treatment response and high mortality rate in patients. High IFP in tumors is driven by three key factors: (i) leaky and immature tumor vasculature; (ii) fibrosis and high density of collagen fibers; and (iii) dense cellular growth.

[0004] The immature and disorganized vasculature of tumors results in increased permeability of vessels. Termed the Enhanced Permeability and Retention (EPR) effect, the EPR effect allows preferential penetration and accumulation of nanoparticles within tumors. Unlike small molecule drugs, nanoparticles can provide the added benefit of longer half-life and remain in circulation for hours or days, which led to the development of multiple nanoparticle formulation of drugs (e.g., Doxil (doxirubicin), Caelyx (doxirubicin), Abraxane (paclitaxel ) etc.) and the use of nanoparticles as imaging agents (e.g., silica nanoparticles, liposomes, iron oxide nanoparticles). However, the vasculature may not be homogenous throughout the tumor and therefore affect the uniform deposition of such nanoparticle agents.

[0005] Zr labeled liposomes are an example of a non-invasive quantitative positron emission tomography (PET) nanoreporter technology that allows personalized therapeutic outcome prediction. Moreover, Zr labeled liposomes can serve as dual-labeled liposomes and provide use as diagnostic tools, e.g., to screen individual subjects for nanotherapy amenability and biodistribution. For example, a stable liposome platform can be efficiently labeled with the radioisotope ⁸⁹Zr and a fluorophore, such as a Cy5 analog or a Cy7 analog.

[0006] Zr labeled liposomes accumulate in vascularized tumor areas via the EPR effect and can be used as companion imaging agents to stratify patients into their appropriate treatment groups. For example, use of ⁸⁹Zr-NRep PET imaging revealed remarkable Doxil accumulation heterogeneity independent of tumor size. Additional details on Zr labeled liposomes are described in U.S. Publication No.20150343100A1, and International Publication No. WO

2015/183876, the disclosures of which are incorporated by reference herein in their entireties.

[0007] Despite the potential advantages of nanoparticles, however, it has been demonstrated that drugs in nanoparticle formulation do not necessarily improve tumor response. Despite their longer bioavailability, nanoparticle formulations of drugs do not demonstrate higher penetration or homogeneous distribution within tumors, negating their potential benefits. Further, currently, there is no imaging modality that can map the uptake pattern of drugs in nanoparticle form and/or prognosticate treatment response from use of such drugs.

[0008] Table 1 outlines currently used tools for increasing perfusion and permeability of dense tumor tissue, including modalities, outcome, therapeutic window, current status, advantages, and shortcomings.

[0009] Therefore, there remains a need for a platform that can locally deliver a drug therapeutic to dense tumor tissues. Summary of invention

[0010] Described herein are systems and methods that combine nanoparticle drugs with electric pulses for enhanced nanoparticle drug delivery to tumors, more particularly to certain “dense” tumors (e.g., pancreatic, colorectal, breast cancers). The systems and methods described herein satisfy the unmet need for a theranostic product for oncology diagnosis and treatment, especially when combined with liposome-delivered drugs. For example, the systems and methods described herein are pertinent to any ductal cancer that has low diffusion coefficient and/or is non responsive to chemotherapy.

[0011] In certain embodiments, devices described herein can be used to apply pulsed electric fields to tumors or other tissue types. Without wishing to be bound by theory, in certain embodiments, pulsed electric fields can alter the interstitial fluid pressure of tumors, increase vascular permeability and flow, cause fluid redistribution within the tumor that leads to reduced collagen density, and increase the interstitial space between cells in the tumor.

[0012] Thus, in certain embodiments, systems, methods, and devices are described herein for the enhanced uptake of administered liposomal or other nanoparticle delivered drugs into tissue of interest/concern, particularly along or in the vicinity of interior lumens of the body. Devices are described herein with an expandable, self-adjusting element that enables

introduction into the lumen in a collapsed form, expansion to provide a plurality of contact points of an electrode/electrode(s) of the device against the inside wall of the lumen, and self- adjustment of the element to maintain contact of the electrode/electrode(s) of the device against the inside wall as it is drawn through the lumen, even as the circumference, diameter, and/or shape of the lumen changes along the length. This provides an improvement over basket catheters that must be sized for use in a specific lumen. [0013] In certain embodiments, the electric pulses delivered to the lumen wall via the electrode(s) of this device increase the total amount of nanoparticle-delivered drugs that enter the tissue of interest, facilitate faster clearance of the nanoparticles from the system, and/or increase homogeneous distribution of the drug throughout the tumor.

[0014] In certain embodiments, devices described herein are used for the delivery of square wave pulses to luminal organs in a circumferential or focal fashion. The square wave pulses can be used to either ablate tumors or other undesirable normal tissue within the organ through irreversible electroporation or nanoporation. The square pulses can also facilitate reversible electroporation of the lumen wall allowing transfection of genetic material or drugs to constituent cells.

[0015] In other embodiments, the devices described herein can be used to deliver high frequency electrical energy into the inner walls of luminal organs. This functions similarly to electrocautery and allows the rapid coagulation of any bleeding in the organ. This is valuable, for example, in the control of internal hemorrhaging, the treatment of bleeding ulcers, and the treatment of diseases such as varices.

[0016] In other manifestations, the devices described herein can be used to deliver radiofrequency energy within hollow organs to partially ablate the lumen wall. This finds application in controlling diseases marked by the hypertrophy of smooth muscle or muscularis of luminal organs. Examples include treatment of asthma, esophageal strictures, and debulking of vascular stenosis.

[0017] In one aspect, the invention is directed to a method of enhancing uptake of an administered composition into a tissue of interest, the method comprising: administering to a subject a therapeutic and/or diagnostic agent; and delivering electric energy (e.g., one or more electric pulses) to an interior surface of a body lumen of the subject (e.g., at one or more points/positions about a circumference of the lumen), thereby enhancing uptake of the administered therapeutic and/or diagnostic agent into the tissue of interest.

[0018] In certain embodiments, the tissue of interest is within, on, and/or in the vicinity of the interior surface of the body lumen of the subject.

[0019] In certain embodiments, the body lumen is the interior of a vessel (e.g., an artery, vein, gastrointestinal tract, esophagus, alimentary canal, respiratory tract, nasal passage, bronchi in the lungs, renal tubule, urinary collecting duct, female genital tract, pathway of the vagina, uterus, fallopian tube, the stomach, sinus tract, biliary duct, pancreatic duct, breast duct, and/or the abdominal cavity).

[0020] In certain embodiments, the tissue is a manifestation of neoplastic disease.

[0021] In certain embodiments, the neoplastic disease is cancer.

[0022] In certain embodiments, the tissue of interest is a dense tumor. Tumor density can be ascertained by pathology (excess collagen or extracellular matrix), imaging (diffusion coefficient across the tumor with MRI), or IFP measurements.

[0023] In certain embodiments, the therapeutic and/or diagnostic agent comprises a nanoparticle.

[0024] In certain embodiments, the therapeutic and/or diagnostic agent comprises a liposome.

[0025] In certain embodiments, the electric energy is delivered before administration of the therapeutic agent.

[0026] In certain embodiments, the electric energy is delivered after administration of the therapeutic agent. [0027] In certain embodiments, the electric energy is delivered using a device as described herein.

[0028] In certain embodiments, the electric energy is delivered by one or more electrodes in the form of one or more electric pulses, wherein the electric pulses are wave pulses (e.g., square waves, sine waves, step waves, triangle waves, or sawtooth waveforms).

[0029] In certain embodiments, the electric pulses are square wave pulses.

[0030] In certain embodiments, the number of pulses applied at a given position along the lumen (e.g., at a given general location of the electrodes) is between 1 and 1000, between 5 and 500, between 10 and 100, or between 10 and 50.

[0031] In certain embodiments, the pulse frequency is between 0.1 Hz and 20 Hz (e.g., between any two of the following values: 0.1 Hz, 0.2 Hz, 0.5 Hz, 1 Hz, 2 Hz, 3 Hz, 4 Hz, 5 Hz, 10 Hz, and 20 Hz).

[0032] In certain embodiments, the pulse width is between 0.001 µs and 1 s, between 0.01 µs and 100 ms, between 0.1 µs and 10 ms, between 1 µs and 1 ms, or between 10 µs and 0.1 ms.

[0033] In certain embodiments, the pulses have sufficient voltage to induce an electric field between the electrodes, wherein the electric field has a strength of at least 100 V/cm, at least 200 V/cm, at least 300 V/cm, at least 400 V/cm, at least 500 V/cm, at least 600 V/cm, at least 700 V/cm, at least 800 V/cm, at least 900 V/cm, or at least 1000 V/cm.

[0034] In certain embodiments, the voltage applied is between 1 and 1000 V, between 5 and 500 V, between 10 and 100 V, or between 10 and 50 V.

[0035] In certain embodiments, the electrodes are needle electrodes.

[0036] In certain embodiments, the electrodes are in direct contact with tumor tissue. [0037] In certain embodiments, the method comprises delivering the electric pulses using a device as described herein.

[0038] In another aspect, the invention is directed to a method for treating a condition comprising delivering electric square wave pulses to tissue of a luminal organ (e.g., an artery, vein, gastrointestinal tract, esophagus, alimentary canal, respiratory tract, nasal passage, bronchi in the lungs, renal tubule, urinary collecting duct, female genital tract, pathway of the vagina, uterus, fallopian tube, the stomach, and/or the abdominal cavity) or other vessel of a subject in a circumferential or focal fashion.

[0039] In certain embodiments, the square wave pulses ablate tumors or other undesirable tissue within the organ or other vessel.

[0040] In certain embodiments, the method comprises performing irreversible electroporation and/or nanoporation of the tissue.

[0041] In certain embodiments, the method comprises performing reversible

electroporation of the tissue.

[0042] In certain embodiments, the tissue is an interior wall of a lumen.

[0043] In certain embodiments, the method comprises transfecting the tissue with genetic material and/or one or more drugs.

[0044] In certain embodiments, the electric square wave pulses are delivered using a device as described herein.

[0045] In another aspect, the invention is directed to a method for treating a condition comprising delivering high frequency electrical energy into the inner walls of a luminal organ.

[0046] In certain embodiments, the method comprises performing coagulation of bleeding in the luminal organ. [0047] In certain embodiments, the condition is a varix (or varices), internal hemorrhage, or a bleeding ulcer.

[0048] In certain embodiments, the high frequency electrical energy is delivered using a device as described herein.

[0049] In another aspect, the invention is directed to a method for treating a condition comprising delivering radiofrequency energy to a lumen wall of a hollow organ.

[0050] In certain embodiments, the method comprises partial ablation of the lumen wall.

[0051] In certain embodiments, the condition is hypertrophy of smooth muscle or muscularis of luminal organs.

[0052] In certain embodiments, the condition is asthma, esophageal strictures, or vascular stenosis.

[0053] In certain embodiments, the radiofrequency energy is delivered using a device as described herein.

[0054] In another aspect, the invention is directed to a device for treating and/or diagnosing a condition in a subject, the device comprising a catheter comprising an expandable element at a distal end that maintains contact with the interior surface of a body lumen at at least two points about a circumference of the body lumen, the expandable element capable of delivering electric energy (e.g., one or more electric pulses) at the at least two points.

[0055] In certain embodiments, the expandable element has an adjustable diameter such that the expandable element is capable of changing diameter (increasing and/or decreasing diameter) as the catheter is drawn along a length of the body lumen, so as to maintain contact between the body lumen at at least two points as the catheter is drawn along the length of the body lumen. [0056] In certain embodiments, the expandable element comprises an electrically conducting material.

[0057] In certain embodiments, the expandable element is disposed concentrically around substantially the (e.g., entire) outer circumference of the catheter.

[0058] In certain embodiments, the expandable element is disposed around a fraction of the circumference of the catheter.

[0059] In certain embodiments, the expandable element is a basket.

[0060] In certain embodiments, the device comprises a second expandable element.

[0061] In certain embodiments, the device comprises a handle, wherein the handle can be manipulated to cause the expandable element to expand from a first, compressed state to a second, expanded state.

[0062] In certain embodiments, the expandable element has a fully expanded

circumference at its widest of at least approximately 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm, 8 cm, 9 cm, 10 cm, 11 cm, 12 cm, 13 cm, 14 cm, 15 cm, 16 cm, 17 cm, 18 cm, 19 cm, 20 cm, 22 cm, 24 cm, 26 cm, 28 cm, 30 cm or more.

[0063] In certain embodiments, the expandable element has a fully expanded

circumference at its widest of between 1 cm and 100 cm, of between 3 cm and 75 cm, of between 5 cm and 50 cm, of between, 7 cm and 30 cm, or between 10 cm and 20 cm.

[0064] In certain embodiments, the expandable element has an adjustable diameter or an adjustable shape after expansion within the body lumen (e.g., in certain embodiments, the expandable element is capable of decreasing its diameter by at least approximately 2%, 5%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 99% of its maximum diameter while still maintaining contact with the interior surface of a lumen of varying diameter, e.g., as the element is drawn through the lumen).

[0065] In another aspect, the invention is directed to a composition comprising a nanoparticle for use in a method for treating a neoplastic disease in a subject, wherein the method comprises: administering to a subject having a tissue of interest the nanoparticle; and delivering one or more electric pulses to an interior surface of a body lumen of the subject, thereby enhancing uptake of the administered therapeutic and/or diagnostic agent into the tissue of interest.

[0066] In another aspect, the invention is directed to a composition comprising a nanoparticle comprising a radiolabel for use in a method of in vivo diagnosis of a neoplastic disease in a subject, wherein the method comprises: administering to a subject having a tissue of interest the nanoparticle; and delivering one or more electric pulses to an interior surface of a body lumen of the subject, thereby enhancing uptake of the administered therapeutic and/or diagnostic agent into the tissue of interest.

[0067] In certain embodiments, the nanoparticle is a liposome.

[0068] In certain embodiments, the electric pulses are delivered before administration of the nanoparticle.

[0069] In certain embodiments, the electric pulses are delivered after administration of the nanoparticle.

[0070] Elements of the embodiments involving one aspect of the invention (e.g., methods) can be applied in embodiments involving other aspects of the invention (e.g., devices), and vice versa. Definitions

[0071] In order for the present disclosure to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification.

[0072] In this application, the use of "or" means "and/or" unless stated otherwise. As used in this application, the term "comprise" and variations of the term, such as "comprising" and "comprises," are not intended to exclude other additives, components, integers or steps. As used in this application, the terms "about" and "approximately" are used as equivalents. Any numerals used in this application with or without about/approximately are meant to cover any normal fluctuations appreciated by one of ordinary skill in the relevant art. In certain

embodiments, the term "approximately" or "about" refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0073] “Administration”: The term "administration" refers to introducing a substance into a subject. In general, any route of administration may be utilized including, for example, parenteral (e.g., intravenous), oral, topical, subcutaneous, peritoneal, intraarterial, inhalation, vaginal, rectal, nasal, introduction into the cerebrospinal fluid, or instillation into body compartments. In some embodiments, administration is oral. Additionally or alternatively, in some embodiments, administration is parenteral. In some embodiments, administration is intravenous. [0074] “Associated”: As used herein, the term“associated” typically refers to two or more entities in physical proximity with one another, either directly or indirectly (e.g., via one or more additional entities that serve as a linking agent), to form a structure that is sufficiently stable so that the entities remain in physical proximity under relevant conditions, e.g., physiological conditions. In some embodiments, associated moieties are covalently linked to one another. In some embodiments, associated entities are non-covalently linked. In some embodiments, associated entities are linked to one another by specific non-covalent interactions (e.g., by interactions between interacting ligands that discriminate between their interaction partner and other entities present in the context of use, such as, for example. streptavidin/avidin interactions, antibody/antigen interactions, etc.). Alternatively or additionally, a sufficient number of weaker non-covalent interactions can provide sufficient stability for moieties to remain associated. Exemplary non-covalent interactions include, but are not limited to, electrostatic interactions, hydrogen bonding, affinity, metal coordination, physical adsorption, host-guest interactions, hydrophobic interactions, pi stacking interactions, van der Waals interactions, magnetic interactions, electrostatic interactions, dipole-dipole interactions, etc.

[0075] “Substantially”: As used herein, the term“substantially”, and grammatic equivalents, refer to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the art will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result.

[0076] “Subject”: As used herein, the term“subject” includes humans and mammals (e.g., mice, rats, pigs, cats, dogs, and horses). In many embodiments, subjects are be mammals, particularly primates, especially humans. In some embodiments, subjects are livestock such as cattle, sheep, goats, cows, swine, and the like; poultry such as chickens, ducks, geese, turkeys, and the like; and domesticated animals particularly pets such as dogs and cats. In some embodiments (e.g., particularly in research contexts) subject mammals will be , for example, rodents (e.g., mice, rats, hamsters), rabbits, primates, or swine such as inbred pigs and the like.

[0077] “Therapeutic agent”: As used herein, the phrase "therapeutic agent" refers to any agent that has a therapeutic effect and/or elicits a desired biological and/or pharmacological effect, when administered to a subject.

[0078] "Treatment": As used herein, the term "treatment" (also "treat" or "treating") refers to any administration of a substance that partially or completely alleviates, ameliorates, relives, inhibits, delays onset of, reduces severity of, and/or reduces incidence of one or more symptoms, features, and/or causes of a particular disease, disorder, and/or condition. Such treatment may be of a subject who does not exhibit signs of the relevant disease, disorder and/or condition and/or of a subject who exhibits only early signs of the disease, disorder, and/or condition. Alternatively or additionally, such treatment may be of a subject who exhibits one or more established signs of the relevant disease, disorder and/or condition. In some embodiments, treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, and/or condition. In some embodiments, treatment may be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of development of the relevant disease, disorder, and/or condition.

[0079] Drawings are presented herein for illustration purposes, not for limitation. Brief description of drawings

[0080] The foregoing and other objects, aspects, features, and advantages of the present disclosure will become more apparent and better understood by referring to the following description taken in conduction with the accompanying drawings, in which:

[0081] FIG.1 shows a catheter according to one embodiment with a basket extended to maximum size.

[0082] FIG.2A-D shows a catheter according to one embodiment with a handle that can control the diameter to which the basket unfurls.

[0083] FIG.3 shows a catheter according to one embodiment with a basket that is unfurled to make contact within the right main bronchus in swine.

[0084] FIG.4 shows exemplary results with CT images post-energy delivery (B) and after withdrawal of the catheter (A). The arrow points to the unfurled electrode making contact with the bronchial wall. The electrodes can be seen as small white dots. After energy delivery there is congestion, and an inflammation region surrounding the bronchus with a hyperintense appearance. There is no evidence of air leak (perforation) which is a common serious complication after energy delivery in the lung.

[0085] FIG.5 shows a cross-sectional view of the lung (cut perpendicular to the bronchus) with an exemplary location of the basket placement within the bronchus matching hyperemic regions (arrow, B). There is a region of hemorrhage and congestion surrounding the bronchus, extending 3-4 cm into parnechyma on each side. A cross section of the bronchus (cut along the bronchus) shows red lines on the internal surface and uniform effects similar to those seen in the perpendicular view (arrow, A). [0086] FIG.6A shows a catheter according to one embodiment wherein a first expandable element is a basket arranged in a planar fashion. FIG.6B shows a catheter according to one embodiment, further comprising a balloon configured such that inflating the balloon would cause the basket to encapsulate a tumor infiltrating the lumen.

[0087] FIG.7 shows a schematic of bilateral MiaPaCa2 tumors in a rodent model. The right flank tumor was treated with electric pulses, the left flank tumor was not treated.

[0088] FIG.8A-F shows autoradiography results of Cohort 1, which was first injected with a drug and then treated with pulsed electric fields described herein, compared to controls that were not treated with pulsed electric fields.

[0089] FIG.9 shows relative radiation from the autoradiography results from Cohort 1, where tumors were injected with drug and then treated with pulses of electric field, compared to controls (i.e., tumors not receiving any drug). Results suggest that pulsed electric fields increase uptake by tumors.

[0090] FIG.10 shows relative radiation from the autoradiography results from Cohort 2, where the tumors treated with pulses of electric field and then injected with a drug, compared to controls (i.e., tumors not receiving any drug). Results suggest that pulsed electric fields increase uptake by tumors.

[0091] FIG.11 shows PET imaging of Cohort 1 (e.g., mouse 1 (M1), mouse 2 (M2)), and control. The top row shows animals 2 hours post pulse delivery, and the bottom row shows animals at 24 hours post pulse delivery. The PET images display differences between the tumors injected with drug and then treated with pulses of electric field compared to controls (i.e., tumors not receiving any drug). [0092] FIG.12 shows PET imaging of Cohort 2 (e.g., mouse 3 (M3), mouse 4 (M4)), and control. The top row shows animals 2 hours post pulse delivery, and the bottom row shows animals at 24 hours post pulse delivery. The PET images display differences between the tumors treated with pulses of electric field and then injected with a drug compared to controls (i.e., tumors not receiving any drug).

[0093] FIGS.13A-13C show PET imaging of Cohorts 1 and 2 imaged after 24 hours.

[0094] FIG.14 shows a schematic describing the secondary effects of electric pulses on tumor microvasculature and connective tissue (B), compared to untreated tumors (A). These effects are expected to increase the extravasation and retention of liposomal nanoparticles in treated tumors.

[0095] FIG.15A-F shows the uptake of radiotracer and drug in treated and untreated tumors in mice bearing unilateral tumors, measured at different time points. Asterisk indicates data points which were more than 3 quartiles from the mean (outliers).

[0096] FIG.16 shows a comparison of rate of uptake (compared to maximum uptake at 48 hours when nanoparticles clear completely from blood circulation) of tracer (A) and drug (B) in treated and untreated tumors in mice bearing unilateral tumors. Correlation is shown of tracer and drug uptake in treated (C) and untreated (D) tumors combining data from all time points at which measurements were performed.

[0097] FIG.17 shows a comparison of tracer and drug with tumor weight in treated (A- C) and untreated (D-F) tumors.

[0098] FIG.18 shows injection of radiotracer before pulse delivery demonstrating uptake at the 2 hour time point (A). Similar uptake cannot be seen when the tracer was injected one hour following pulse delivery (B). However, 24 hours following treatment, there was no appreciable impact of injection/treatment on order and uptake (C and D). Solid arrows indicate tumors receiving treatment, and dashed arrows indicate contralateral untreated tumors.

[0099] FIG.19 shows (A) an isosurface representing tracer uptake in a treated tumor (solid arrow). The untreated tumor (dashed arrow) cannot be adequately visualized. Liver (arrowhead) and spleen (diamond) can also be seen in the image. Autoradiography of tumors treated with electric pulses (B) demonstrates even distribution of tracer everywhere in the tumor, while similar measurements performed on untreated tumors (C) suggest pockets of tracer deposition.

[0100] FIG.20 shows (A) autoradiography of tumors treated with electric pulses demonstrating distribution of tracer throughout the tumor. Graph (B) depicts radiation counts comparing treated tumors with contralateral control.

[0101] FIG.21 shows a low magnification image (scale bar = 1 mm) showing both needle tracts (arrows) (A). The box insert in (A) is shown in higher magnification in (B), displaying a region of necrosis (dashed boundary) closely bounded by the defect in the tumor from needle placement (arrow). Scale bar = 0.5 mm.

[0102] FIG.22 shows an embodiment of the device: (a) Two looped electrodes (arrows) are buried in a 9Fr catheter. Electrodes are placed at a right angle. (b) The electrode can be expanded and contact the bronchial wall by pushing it through the catheter.

[0103] FIG.23 shows radio images of a sample procedure. (a) An expandable catheter electrode (black arrow) was placed in the bronchi through an intubation tube under fluoroscopy. (b) After expanding the electrodes, CT shows that electrodes are in contact with the bronchial wall (white arrows). (c) Dense consolidation (white arrowheads) appeared after ablation. [0104] FIG.24 shows specimen of untreated and treated bronci. (a) Depicted here is the internal surface of the untreated bronchus opened along the way of the main duct. (b) Depicted here is the internal surface of the treated bronchus. Compared to the untreated bronchus, red lines are seen on the internal surface of treated bronchus (white arrowhead). (c) Depicted here is a lung cut perpendicular to the non-treated bronchus. (d) Depicted here is a lung cut

perpendicular to the treated bronchus. Color of the lung tissue surrounding the treated bronchus changed to dark red.

[0105] FIG.25 shows H&E stained tissue specimens. (a) Untreated bronchus (1.25x); (b) Untreated bronchus (20x); (c) Untreated lung parenchyma (20x); (d) Treated bronchus (1.25x); (e) Treated lung parenchyma (20x); (f) Treated bronchus (20x). Compared to untreated tissue (b, c), sloughing of bronchial epithelium (black arrows), hemorrhage (black arrowheads), hyperemic congestion (white arrowheads), and necrosis of submucosal glands (white arrows) are seen in treated tissue (e). At surrounding lung parenchyma (d), inflammatory cell infiltration and hemorrhage in alveolar space can be seen (yellow arrowheads). In addition, interlobular septum became thicker due to edema (yellow arrows).

[0106] FIG.26 shows Simulation images. Simulation of catheter directed endoluminal IRE: Simulation of IRE with placing 2 electrodes along the bronchus (B: Bronchus, V: Vessel). In both treatments, temperature increases around the electrodes (white arrows). The lesion of preferential passage of electric field into the parenchyma (black arrows) is slightly larger in endoluminal IRE (a) than IRE with 2 electrodes placed around the bronchus (b). Detailed Description

[0107] Throughout the description, where compositions are described as having, including, or comprising specific components, or where methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are

compositions of the present invention that consist essentially of, or consist of, the recited components, and that there are methods according to the present invention that consist essentially of, or consist of, the recited processing steps.

[0108] It should be understood that the order of steps or order for performing certain actions is immaterial so long as the invention remains operable. Moreover, two or more steps or actions may be conducted simultaneously.

[0109] The mention herein of any publication, for example, in the Background section, is not an admission that the publication serves as prior art with respect to any of the claims presented herein. The Background section is presented for purposes of clarity and is not meant as a description of prior art with respect to any claim.

[0110] Described herein are methods and devices for electroporation. Square wave electric pulses are known to induce pore formation in the cell membrane, and this phenomenon is termed electroporation. Electroporation is commonly used for the transfection of cells with drugs and genetic material that normally cannot permeate an intact cell membrane.

Electroporation has also been used for the image-guided ablation of colorectal metastases, pancreatic cancer and renal tumors in patients (See Narayanan et. al., Cardiovasc Intervent Radiol.2014 Dec;37(6):1523-9.; Dollinger M J et. al., Vasc Interv Radiol.2014

Oct;25(10):1589-94; Scheffer HJ et. al. J Vasc Interv Radiol.2014 Jul;25(7):997-1011). Apart from permeabilization of the cell membrane, the application of electric pulses can also cause alterations of the vasculature and blood flow in the treated tissue. Using an ⁸⁶RbCl radiotracer, it has been demonstrated that treatment of tumors with low voltage electric pulses can transiently increase blood flow, with limited cytotoxic effects (See Sersa G, et. al., Eur J Cancer.1999 Apr;35(4):672-7). In one example, the changes in vascular flow were observed to normalize by 24 hours after treatment. Other examples used in vivo optical imaging and a dorsal skinfold window chamber model to study the effect of electric pulses on normal vasculature using a 2000KDa FITC labeled Dextran (See Markelc B, et al., J Membr Biol.2012 Sep;245(9):545-54. The results suggest that electric pulses can cause pronounced vasostriction of arterioles that resolves within 30-60 minutes of pulse delivery. Treatment with electric pulses can also increase permeability of these vessels, causing extravasation of fluorescent dye into the surrounding tissue. Rounding of endothelium can be observed on histology analysis, with inflammation and edema in the tissues adjacent to the core treatment area. In another example, a delay was observed between delivery and tissue permeation of high molecular weight dyes, which was not observed in the low molecular weight version of the same dye (See Bellard E, et. al., J Control Release.2012 Nov 10;163(3):396-403.) In that instant, the permeability of blood vessels was found to gradually normalize post-treatment but did not recover fully during the experimental timeframe (one hour). These prior studies clearly demonstrate that treatment with electric pulses can cause alterations to blood vessels that persist longer than the effect of electroporation itself, where the cells are in a permeable state for only minutes.

[0111] Described herein are systems and methods for manipulation of tumors with pulsed electric fields. Without wishing to be bound by theory, techniques and treatment parameters were identified for the application of pulsed electric fields to alter the interstitial fluid pressure of tumors. In certain embodiments, devices such as those described herein can be used to apply pulsed electric fields to tumors. Without wishing to be bound by theory, in certain embodiments, pulsed electric fields can increase vascular permeability and flow, causing fluid redistribution within the tumor that leads to reduced collagen density, and can increase the interstitial space between cells in the tumor.

[0112] Pulsed electric fields can be applied as a series of low voltage wave pulses. The wave pulses can be square waves, sine waves, step waves, triangle waves, or have sawtooth waveforms. The number of wave pulses delivered at a given position can be 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more pulses, for example. Electric pulses can be applied in quantities of between 1 and 1000, between 5 and 500, between 10 and 100, or between 10 and 50, for example. The pulses can have a frequency of 0.1 Hz, 0.2 Hz, 0.5 Hz, 1 Hz, 2 Hz, 3 Hz, 4 Hz, 5 Hz, 10 Hz, or 20 Hz. The pulse width can be between 0.001 µs and 1 s, between 0.01 µs and 100 ms, between 0.1 µs and 10 ms, between 1 µs and 1 ms, or between 10 µs and 0.1 ms.

[0113] The pulses can have sufficient voltage to induce an electric field between the electrodes, wherein the electric field has a strength of at least 100 V/cm, 200 V/cm, 300 V/cm, 400 V/cm, 500 V/cm, 600 V/cm, 700 V/cm, 800 V/cm, 900 V/cm, or 1000 V/cm. The voltages applied can be between 1 and 1000 V, between 5 and 500 V, between 10 and 100 V, or between 10 and 50 V.

[0114] For example, in one embodiment, the method described herein is a treatment that applies a series (10-50) of low voltage (less than 1000 V) square wave DC pulses of 1 µs to 1 ms pulse width. In certain embodiments, such a treatment can be rapid, lasting less than a minute, and the effects on the tumor can start manifesting a few hours (4 - 8 hours) following treatment. In one embodiment, the effects can persist for a period of up to about 57 days. In certain embodiments, tumors are treated with pulses with sufficient voltage to induce an electric field with strength of 700V/cm between the needle electrodes, with 10 pulses delivered at 1 Hz with a pulse length of 90 microseconds.

[0115] The pulses can be applied to tissues, such as tumors, using either pairs of needle electrodes or pairs of plate electrodes for transcutaneous noninvasive treatment. Electrodes may be configured as required by tissue type or application. In certain embodiments, pulses may be applied using 2, 3, 4, 5, 6, 7, 8, or more electrodes.

[0116] Described herein are devices and methods for delivering therapeutic or diagnostic agents to a tissue. In certain embodiments, the devices and methods include electroporation mediated vascular changes and edema, passively modifying the tumor microenvironment, resulting in changes to its permeability and retention properties. In certain embodiments, the devices and methods include causing an Enhanced Permeability and Retention (EPR) effect in the tissue. In certain embodiments, the devices and methods include causing selective uptake of nanoparticles in tumors. In certain embodiments, the devices and methods include treating tumors with electric pulses. In certain embodiments, the devices and methods can cause the uptake of nanoparticles. In certain embodiments, the devices and methods can cause an uptake of nanoparticles by a tumor, wherein the uptake by the tumor treated with electric pulses is increased compared to the uptake by a tumor not treated with electric pulses. In certain embodiments, uptake is independent of electropermeabilization of the cell membrane. In certain embodiments, the uptake of nanoparticles by the tumor treated with electric pulses is increased compared to the uptake by a tumor not treated with electric pulses, and this increase in uptake is independent of electropermeabilization of the cell membrane. [0117] Also described herein are systems and methods for mapping pulsed electric field tumors with nanoparticles and predicting therapy response in subjects (e.g., mice) with bilateral tumors.

Imaging agents

[0118] In certain embodiments, the compositions described herein include (i) imaging agents that are, or are associated with, the therapeutic agent, and/or (ii) imaging agents that are associated with, or are a part of, liposomes or other nanoparticle-based delivery platform, e.g., Zr labeled liposomes. Zr labeled liposomes that can be used with the methods and devices described herein are described in U.S. Publication No.20150343100A1, and International Publication No. WO 2015/183876, the disclosures of which are incorporated by reference herein in their entireties. In certain embodiments, the imaging agents can include radiolabels, radionuclides, radioisotopes, fluorophores, fluorochromes, dyes, metal lanthanides, paramagnetic metal ions, superparamagnetic metal oxides, ultrasound reporters, x-ray reporters, and/or fluorescent proteins.

[0119] In certain embodiments, radiolabels comprise ^99mTc, ¹¹¹In, ⁶⁴Cu, ⁶⁷Ga, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁵³Sm, ¹⁷⁷Lu, ⁶⁷Cu, ¹²³I, ¹²⁴I, ¹²⁵I, ¹¹C, ¹3N, ¹⁵O, ¹⁸F,¹⁸⁶Re, ¹⁸⁸Re, ¹⁵³Sm, ¹⁶⁶Ho, ¹⁷⁷Lu, ¹⁴⁹Pm, ⁹⁰Y, ²¹²Bi, ¹⁰³Pd, ¹⁰⁹Pd, ¹⁵⁹Gd, ¹⁴⁰La, ¹⁹⁸Au, ¹⁹⁹Au, ¹⁶⁹Yb, ¹⁷⁵Yb, ¹⁶⁵Dy, ¹⁶⁶Dy, ⁶⁷Cu, ¹⁰⁵Rh, ¹¹¹Ag, ⁸⁹Zr, and ¹⁹²Ir. In some embodiments, paramagnetic metal ions comprise Gd(III), Dy(III), Fe(III), and Mn(II). In some embodiments, Gadolinium (III) contrast agents comprise Dotarem, Gadavist, Magnevist, Omniscan, OptiMARK, and Prohance. In certain embodiments, x-ray reporters comprise iodinated organic molecules or chelates of heavy metal ions of atomic numbers 57 to 83. [0120] In certain embodiments, PET (Positron Emission Tomography) tracers are used as imaging agents. In some embodiments, PET tracers comprise ⁸⁹Zr, ⁶⁴Cu, [¹⁸F]

fluorodeoxyglucose.

[0121] In certain embodiments, fluorophores comprise fluorochromes, fluorochrome quencher molecules, any organic or inorganic dyes, metal chelates, or any fluorescent enzyme substrates, including protease activatable enzyme substrates. In some embodiments,

fluorophores comprise long chain carbophilic cyanines. In other embodiments, fluorophores comprise DiI, DiR, DiD, and the like. Fluorochromes comprise far red, and near infrared fluorochromes (NIRF). Fluorochromes include but are not limited to a carbocyanine and indocyanine fluorochromes. In some embodiments, imaging agents comprise commercially available fluorochromes including, but not limited to Cy5.5, Cy5 and Cy7 (GE Healthcare); AlexaFlour660, AlexaFlour680, AlexaFluor750, and AlexaFluor790 (Invitrogen); VivoTag680, VivoTag-S680, and VivoTag-S750 (VisEn Medical); Dy677, Dy682, Dy752 and Dy780

(Dyomics); DyLight547, DyLight647 (Pierce); HiLyte Fluor 647, HiLyte Fluor 680, and HiLyte Fluor 750 (AnaSpec); IRDye 800CW, IRDye 800RS, and IRDye 700DX (Li-Cor); and

ADS780WS, ADS830WS, and ADS832WS (American Dye Source) and Kodak X-SIGHT 650, Kodak X-SIGHT 691, Kodak X-SIGHT 751 (Carestream Health).

Therapeutic Agents

[0122] In certain embodiments, the compositions described herein include a therapeutic agent. In certain embodiments, the therapeutic agent is associated with a liposome or other nanoparticle-based delivery platform, e.g., Zr labeled liposome, which is administered to the subject. In certain embodiments, the therapeutic agent is an anti-cancer agent. In certain embodiments, the anti-cancer agent is a chemotherapeutic agent. For example, in certain embodiments, the therapeutic agent is an alkylating agent, an antimetabolite, an anthracycline, an antibiotic, and camptothecin, a vince alkaloid, a taxane, a platinum compound, a hormonal agent, a cytotoxic agent, an enzyme, a microtubule damaging agent, a topoisomerase-1 inhibitor, a topoisomerase-2 inhibitor, a tyrosine proteinkinase inhibitor, an EGF receptor inhibitor, an angiogenesis inhibitor, a protease inhibitor, a glucocorticoid, an estrogen, an aromatase inhibitor, an antiandrogen, a 5-alpha inhibitor, a GnRH analog, or a progestin.

Administration

[0123] Pharmaceutical compositions incorporating the nanoparticle described herein may be administered according to any appropriate route and regimen. In some embodiments, a route or regimen is one that has been correlated with a positive therapeutic benefit.

[0124] In certain embodiments, the exact amount administered may vary from subject to subject, depending on one or more factors as is well known in the medical arts. Such factors may include, for example, one or more of species, age, general condition of the subject, the particular composition to be administered, its mode of administration, its mode of activity, the severity of disease; the activity of the specific nanoparticle employed; the specific

pharmaceutical composition administered; the half-life of the composition after administration; the age, body weight, general health, sex, and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed and the like. Pharmaceutical compositions may be formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions will be decided by an attending physician within the scope of sound medical judgment. [0125] Compositions described herein may be administered by any route, as will be appreciated by those skilled in the art. In certain embodiments, compositions described herein are administered by oral (PO), intravenous (IV), intramuscular (IM), intra-arterial,

intramedullary, intrathecal, subcutaneous (SQ), intraventricular, transdermal, interdermal, intradermal, rectal (PR), vaginal, intraperitoneal (IP), intragastric (IG), topical (e.g., by powders, ointments, creams, gels, lotions, and/or drops), mucosal, intranasal, buccal, enteral, vitreal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; as an oral spray, nasal spray, and/or aerosol, and/or through a portal vein catheter. In certain embodiments, the compositions are administered directly to a tissue via a catheter.

[0126] In some embodiments, the pharmaceutical compositions and/or Zr labeled liposomes thereof may be administered intravenously (e.g., by intravenous infusion), by intramuscular injection, by intratumoural injection, and/or via portal vein catheter, for example. However, the subject matter described herein encompasses the delivery of pharmaceutical compositions and/or Zr labeled liposomes thereof in accordance with embodiments described herein by any appropriate route taking into consideration likely advances in the sciences of drug delivery.

Diseases and Conditions

[0127] In certain embodiments, devices and methods described herein can be used to diagnose and/or treat the diseases and conditions situated within, on, and/or in the vicinity of the interior surface of a body lumen. In certain embodiments, the body lumen is the interior of a vessel, such as the central space in an artery or vein; the interior of the gastrointestinal tract; the interior of the respiratory tract; the pathways of the bronchi in the lungs; the interior of renal tubules and urinary collecting ducts; the pathways of the female genital tract, starting with a single pathway of the vagina, splitting up in two lumina in the uterus, both of which continue through the fallopian tubes; the interior of the stomach; sinus tract; biliary duct; pancreatic duct; breast duct; or the abdominal cavity.

[0128] The diseases and conditions that can be diagnosed and/or treated with the devices and methods described herein include neoplastic disease. In certain embodiments, the neoplastic disease is cancer. In certain embodiments, the cancer is Stage I cancer, Stage II cancer, Stage III cancer, Stage IV cancer, carcinoma, lymphoma, sarcoma, myeloma, blastoma, and

adenocarcinoma, bone cancer, breast cancer, colon/rectum cancer, lung cancer, nasopharyngeal cancer, oral cavity and oropharyngeal cancer, small intestine cancer, stomach cancer, uterine sarcoma, or vaginal cancer.

Devices

[0129] Described herein are devices to administer to a tissue a voltage or an agent, or both, wherein the agent is an imaging agent or a therapeutic agent, or both. In certain

embodiments, the voltage, the imaging agent or therapeutic agent, alone or in combination, are administered systemically. In certain embodiments, the voltage, the imaging agent or therapeutic agent, alone or in combination, are administered locally directly to the tissue to be treated.

[0130] In certain embodiments, the device comprises a catheter. In certain embodiments, the catheter is an electrode catheter. In certain embodiments, the catheter is an electrode catheter, wherein the catheter comprises a needle end, wherein the needle end is an electrode. In certain embodiments, the electrode catheter further comprises one or more additional elements capable of conducting an electric current.

[0131] In certain embodiments, the device comprises one or more electrodes. The electrodes can be linear, flat, round, spherical, cylindrical, square, cubic, triangular, pyramidic, hexagonal, or any combination thereof. The electrodes can be in the form of a needle, a wire, a tine, a hollow tube, a coil, a loop, a sling, a blade, a fork, a spoon, a surface, or a cage.

[0132] In certain embodiments, the device comprises a needle. In certain embodiments, the needle is an electrode. In certain embodiments, the needle is a 7 G, 8 G, 9 G, 10 G, 11 G, 12 G, 13 G, 14 G, 15 G, 16 G, 17 G, 18 G, 19 G, 20 G, 21 G, 22 G, 22s G, 23 G, 24 G, 25 G, 26 G, 26s G, 27 G, 27 G, 28 G, 29 G, 30 G, 31 G, 32 G, 33 G, or 34 G needle.

[0133] In certain embodiments, the device comprises one or more expandable elements, e.g., a basket, a stent, or a balloon. In certain embodiments, the expandable element is capable of conducting an electric current. In certain embodiments, the expandable element comprises an electrically conducting material. In certain embodiments, the expandable element is a wire element. In certain embodiments, the catheter comprises a needle end and an expandable element, wherein the needle end and the expandable element are moveable independently from each other. In certain embodiments, the expandable element, when in its constrained or un- expanded state, has a sharp or pointed end that may be inserted into the tissue. In certain embodiments, the expandable element, when in its constrained or un-expanded state, causes the catheter to have a total length greater than its length with an expandable element in the expanded state. The expandable element may maintain a collapsed configuration during insertion and/or positioning within the lumen, and then expand when the distal end of the device is positioned where desired within the lumen, thereby achieving contact between electrodes of the expandable element (or electrodes otherwise coupled to the expandable element) and the interior surface of the lumen. In addition to being expandable, in certain embodiments, the expandable element is also capable of automatical adjustment (e.g., via its shape, flexibility, configuration, and the like) to vary its circumference during use. For example, a basket configuration comprising multiple tines is capable of maintaining contact with the interior surface of a lumen as the device is drawn along the length of the lumen, where the lumen has varying internal diameter.

[0134] In certain embodiments, the device comprises a basket (See FIG.1). In certain embodiments, the basket is comprised of 4 tines. In certain embodiments, the basket is comprised of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 tines, or about 25 tines, about 30 tines, about 35 tines, about 40 tines, about 45 tines, or about 50 or more tines. In certain embodiments, the expandable element is configured to deliver electrical energy into the walls of hollow organs. Hollow organs include the bronchus, esophagus, urinary tract, intestine, stomach, abdominal cavity, bladder, blood vessel, heart, and lung. In certain embodiments, the expandable element is configured to deliver electrical energy into a cavity or lumen of any size or shape without loss of contact. In certain embodiments, the expandable element has the basic shape of a sphere, a cylinder, a cone, a truncated cones, a cube, a prisms, or a pyramid, or a combination thereof. In certain embodiments, the expandable element has a fully expanded circumference at its widest of approximately 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm, 8 cm, 9 cm, 10 cm, 11 cm, 12 cm, 13 cm, 14 cm, 15 cm, 16 cm, 17 cm, 18 cm, 19 cm, 20 cm, 22 cm, 24 cm, 26 cm, 28 cm, 30 cm or more. In certain embodiments, the expandable element has a fully expanded circumference at its widest of between 1 cm and 100 cm, of between 3 cm and 75 cm, of between 5 cm and 50 cm, of between, 7 cm and 30 cm, or between 10 cm and 20 cm. In certain embodiment, the expandable element has an adjustable diameter or an adjustable shape when fully expanded. In certain embodiments the expandable element is capable of decreasing its diameter by approximately 2%, 5%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 99% of its maximum diameter while still maintaining contact with the interior surface of a lumen of varying diameter, e.g., as the element is drawn through the lumen). In certain embodiments, the expandable element has a length when fully expanded of about 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm, 8 cm, 9 cm, 10 cm, 11 cm, 12 cm, 13 cm, 14 cm, 15 cm, 16 cm, 17 cm, 18 cm, 19 cm, 20 cm, 22 cm, 24 cm, 26 cm, 28 cm, 30 cm or more. In certain embodiments, the expandable element is constructed to be compliant, therefore adjusting to the size or shape, or both, of the lumen or other cavity within it is placed.

[0135] In certain embodiments, the electrode catheter comprises an inflatable element. In certain embodiments, the inflatable element is a balloon. In certain embodiments, the inflatable element comprises one or more electrically conducting elements. In certain

embodiments, the balloon comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 electrically conducting elements, or about 25 electrically conducting elements, about 30 electrically conducting elements, about 35 electrically conducting elements, about 40 electrically conducting elements, about 45 electrically conducting elements, or about 50 or more electrically conducting elements. In certain embodiments, the electrically conducting elements can be arranged in various patterns on the surface of the inflatable element, for example in rows, columns, circles, on a rectangular grid, on a triangular grid, on hexagonal grid, or in random configuration. In certain embodiments, the inflatable element is configured to deliver electrical energy into the walls of hollow organs. Hollow organs include the bronchus, esophagus, urinary tract, intestine, stomach, abdominal cavity, bladder, blood vessel, heart, and lung. In certain embodiments, the inflatable element is configured to deliver electrical energy into a cavity or lumen of any size or shape without loss of contact. In certain embodiments, the inflatable element has the basic shape of a sphere, a cylinder, a cone, a truncated cones, a cube, a prisms, or a pyramid, or a combination thereof. In certain embodiments, the inflatable element has a fully expanded circumference at its widest of approximately 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm, 8 cm, 9 cm, 10 cm, 11 cm, 12 cm, 13 cm, 14 cm, 15 cm, 16 cm, 17 cm, 18 cm, 19 cm, 20 cm, 22 cm, 24 cm, 26 cm, 28 cm, 30 cm or more. In certain embodiments, the inflatable element has a fully expanded circumference at its widest of between 1 cm and 100 cm, of between 3 cm and 75 cm, of between 5 cm and 50 cm, of between, 7 cm and 30 cm, or between 10 cm and 20 cm. In certain embodiment, the inflatable element has an adjustable diameter or an adjustable shape when fully expanded. In certain embodiments the inflatable element is capable of decreasing its diameter by approximately 2%, 5%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 99% of its maximum diameter while still maintaining contact with the interior surface of a lumen of varying diameter, e.g., as the element is drawn through the lumen). In certain embodiments, the inflatable element has a length when fully expanded of about 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm, 8 cm, 9 cm, 10 cm, 11 cm, 12 cm, 13 cm, 14 cm, 15 cm, 16 cm, 17 cm, 18 cm, 19 cm, 20 cm, 22 cm, 24 cm, 26 cm, 28 cm, 30 cm or more.

[0136] In certain embodiments, the catheter further comprises a handle. In certain specific embodiments, the handle can move, or can cause other elements of the catheter to move, along the length of the catheter.

[0137] In certain embodiments, the handle is disposed such that manipulation of the handle causes change in the size (e.g., the diameter) of the expandable element. Manipulation includes sliding, twisting, squeezing, folding, pushing, pulling, pressing, or splitting the handle or any component or part thereof.

[0138] In certain specific embodiments, the catheter has a distal end, the distal end comprising a needle end, and a proximal end comprising the moveable handle. In certain embodiments, moving the handle, or a part or component thereof, in a proximal direction causes the size of the expandable element (e.g., a basket) to increase, and moving the handle in a distal direction causes the size of the basket to decrease. In certain embodiments, moving the handle, or a part or component thereof, proximally causes the size of the expandable element (e.g., a basket) to decrease, and moving the handle distally causes the size of the basket to increase. FIG.2A-D shows how moving the handle can control the diameter to which the basket unfurls. FIG.3 shows an embodiment wherein the basket is unfurled to make contact within the right main bronchus in swine.

[0139] The expandable element can deliver electric energy to the tissue. FIG.4 shows exemplary results with CT images post-energy delivery (B) and after withdrawal of the catheter (A). FIG.5B shows a cross-sectional view of the lung (cut perpendicular to the bronchus) with an exemplary location of the basket placement within the bronchus matching hyperemic regions (arrow). There is a region of hemorrhage and congestion surrounding the bronchus, extending 3- 4 cm into parenchyma on each side. A cross section of the bronchus (cut along the bronchus) shows red lines on the internal surface and similar uniform effects similar to those seen in the perpendicular view (arrow, FIG.5A).

[0140] In certain embodiments, the expandable element (e.g., a basket) is disposed concentrically around substantially the entire outer circumference of the catheter. In certain embodiments, the expandable element is disposed around a fraction of the circumference of the catheter. In certain embodiments, the catheter comprises a first and a second expandable element, e.g., a basket, a stent, or a balloon. In certain embodiments, the first expandable element is disposed around a fraction of the circumference of the catheter, and the second expandable element is disposed around another fraction of the circumference of the catheter. In certain specific embodiments, the first expandable element is a basket arranged in a planar fashion as shown in FIG.6A In certain specific embodiments, the second expandable element is a balloon, configured such that inflating said balloon would cause said basket to encapsulate a tumor infiltrating the lumen (FIG.6B).

[0141] In certain embodiments, the device is configured such that it is suitable for use with tumors that may be infiltrating a lumen of a hollow organ.

[0142] Described herein are devices that can be used for tissue ablation. In certain embodiments, the catheter can be used to deliver electrical energy to perform ablation using radiofrequency, irreversible electroporation, electrochemical therapy, EStress or other techniques.

[0143] Described herein are devices that can be used for drug delivery. Without wishing to be bound by theory, in certain embodiments, the energy delivered by the devices described herein can facilitate drug delivery through hyperthermia, electroporation, or the use of electric pulses to modulate tissue vasculature and stroma. Without wishing to be bound by theory, in certain embodiments, the devices deliver pulses to increase or expand space, loosening tumor stroma, which (i) increases total amount of nanoparticles that can be absorbed by the tumor, (ii) facilitate faster clearance of nanoparticles from the system, and (iii) causes homogeneous distribution of nanoparticle throughout the tumor. In certain embodiments, the devices can be used in Oncology, wherein ablation or drug delivery can be performed to nonsurgically destroy tumors that cannot be removed with surgery. In other embodiments, the devices can be used to treat or control nonmalignant disease conditions, including drug or gene delivery for acute lung injury, infections in the lung, and conditions like Asthma or COPD. In other embodiments, the devices can be used to opening the blood-brain barrier for the improved deliver of drugs to the brain. [0144] Described herein is a device that can be used for the delivery of square wave pulses to luminal organs in a circumferential or focal fashion. In certain embodiments, the square wave pulses can be used to ablate either tumors or other undesirable normal tissue within the organ through irreversible electroporation or nanoporation. In certain embodiments, the square pulses can also facilitate reversible electroporation of the lumen wall allowing transfection of genetic material or drugs to constituent cells.

[0145] Described herein is a device that can be used to delivered high frequency electrical energy into the inner walls of luminal organs. In certain embodiments, the device is configured to function similar to electrocautery to allow the rapid coagulation of any bleeding in the organ. In certain embodiments, the device can be used to treat certain conditions such as varices, internal hemorrhage, and/or bleeding ulcers.

[0146] Described herein is a device that can be used to deliver radiofrequency energy within hollow organs. In certain embodiments, the device is configured such that its use pursuant to the methods described herein would cause partial ablation of the lumen wall. In certain embodiments, the device can be used to treat certain conditions marked by the hypertrophy of smooth muscle or muscularis of luminal organs, such as asthma, esophageal strictures, and/or debulking of vascular stenosis.

Examples

Combination of pulsed electric fields and liposome drug delivery in pancreatic tumors

EXAMPLE 1

[0147] The use of radiolabeled nanoparticles allows imaging and quantification of the change or difference in distribution and retention of particles in treated tumors (See International Publication No. WO 2015/183876, attached hereto). Radiolabeled liposomal nanoparticles show a linear correlation in tumor uptake when compared with simultaneously administered liposomal drugs. Therefore, co-administration of a nanoparticle drug formulation with a small dose of radiolabeled liposome can serve as method to map the uptake and relative distribution of therapeutic agents in tumors treated with pulsed electric fields. Subsequently, the uptake value of the radiotracer can be used to establish the concentration of the therapeutic agent at different locations in the tumor, and therefore serve as a tool to prognosticate treatment response at a very early stage.

[0148] In a first study, tumors in one hind limb of a rodent were treated with pulsed electric fields, and a contralateral tumor served as a control. Zr labeled liposomal nanoparticles were injected either before or after electrical treatment.

Methods

[0149] Pulsed electric fields were applied to tumors in mice to determine if an increase in drug uptake via EPR effect occurred. Timing of the injection of drug and electric pulse treatment was also investigated to determine effect on uptake by a tumor. As depicted in the schematic of FIG.7, bilateral MiaPaCa2 tumors in two cohorts were treated with electric pulses (e.g., right flank) or not treated (e.g., left flank). Cohort 1 was first injected with the drug, treated with pulsed electric fields, imaged, and sacrificed to determine level of drug uptake by the tumor. Cohort 2 was first treated with pulsed electric fields, injection of drug, imaged and sacrificed to determine level of drug uptake by the tumor.

[0150] Two 19G electrodes were placed in the tumor as far away from each other as possible. An ECM 835 square wave generator was used to deliver electric pulses. 700 V/cm was applied between the electrodes, 10 pulses at 100 µs at 1 Hz.

Results

[0151] FIG.8A-F shows autoradiography results of Cohort 1, which was first injected with a drug and the treated with pulsed electric fields described above (A-C) compared to controls that were not treated with pulsed electric fields (D-F).

[0152] FIG.9 shows relative radiation from the autoradiography results from Cohort 1 between the tumors injected with drug and then treated with pulses of electric field compared to controls (e.g., tumors not receiving any drug). Results suggest that pulse electric fields increase uptake by tumors.

[0153] FIG.10 shows relative radiation from the autoradiography results from Cohort 2 between the tumors treated with pulses of electric field and then injected with a drug compared to controls (e.g., tumors not receiving any drug). Results suggest that pulse electric fields increase uptake by tumors.

[0154] FIG.11 shows PET imaging of Cohort 1 (e.g., mouse 1 (M1), mouse 2 (M2)), and control. The top row shows animals 2 hours post pulse delivery, and the bottom row shows animals at 24 hours post pulse delivery. The PET images display differences between the tumors injected with drug and then treated with pulses of electric field compared to controls (e.g., tumors not receiving any drug). FIG.12 shows PET imaging of Cohort 2 (e.g., mouse 3 (M4), mouse 4 (M4)), and control. The top row shows animals 2 hours post pulse delivery, and the bottom row shows animals at 24 hours post pulse delivery. The PET images display differences between the tumors treated with pulses of electric field and then injected with a drug compared to controls (e.g., tumors not receiving any drug). Unlike standard electroporation techniques that require the agent to be present in the circulation before pulse delivery, the present technique is agnostic as to when the nanoparticle agent is injected.

[0155] FIGS.13A-13C show Cohorts 1 and 2 imaged after 24 hours. The PET/CT images were processed with a software to extract iso-surfaces corresponding to bone window (white) and the locations positive for radioactivity (gold).

[0156] At 24 hours after treatment, treated tumors were seen to have significantly (1 - 8 fold) higher uptake of the radiolabeled nanoparticle. Autoradiography results suggest that penetration and distribution within the tumor corresponded to the electric field generated within the tumor from application of the pulsed electric field. Autoradiography analysis of control tumors demonstrated non-uniform and non-specific distribution of the liposomes within the tumor.

[0157] Results from these experiments suggest that treatment with pulsed electric fields can alter the tumor to increase the penetration and the amount of nanoparticles. The distribution of nanoparticles seem non-specific to tumor morphology and seem related to the location of the electrodes and the electric field distribution within the treated tissue.

[0158] In summary, the results of using the systems and methods herein showed increased drug uptake in electrically treated tissue (about 2 or more times) compared to controls. These changes were achieved by applying a series (10-50) of low voltage (less than 1000 V) square wave DC pulses of 1 µs to 1 ms pulse width. The treatment can last less than a minute and the effects on the tumor start manifesting a few hours (48 hours) following treatment and can persist up till a period of 57 days.

[0159] Without wishing to be bound to any theory, there is increased uptake of drug via EPR effect in the treated tumor vs. the non-treated tumor. It appears that an increase in uptake of drug is related to the timing of the injection of the drug and treatment. For example, treating post-injection leads to a higher uptake. EXAMPLE 2

[0160] In another experiment, it was studied to what extent electroporation mediated vascular changes and edema passively modified the tumor microenvironment, resulting in changes to its permeability and retention properties (FIG.14). In this experiment a novel ⁸⁹Zr- radiolabeled liposomal nanoparticle (⁸⁹Zr-NRep) was used as an exemplary tracer to monitor the effect of electric pulse treatment on xenograft subcutaneous pancreatic adenocarcinoma tumor model (See Pérez-Medina C, et al., J Nucl Med.2014 Oct;55(10):1706-11. The uptake of ⁸⁹Zr- NRep was compared with the uptake of liposomal doxorubicin as an exemplary drug in treated and untreated tumors.

Materials and Methods

Animal and Tumor Model

[0161] MiaPaca-2 cells were cultured using DME modified to contain 1 mM sodium pyruvate, 1.5 g/L sodium bicarbonate and 4.5 g/L glucose, supplemented with 10% (vol/vol) heat-inactivated FCS and 100 IU penicillin and 100 µg/mL streptomycin.5×106 cells were then injected in athymic nude mice, strain NU(NCr)-Foxn1^nu (Charles River Laboratories,

Wilmington, MA) in 150μl of a 1:1 mixture of growth medium and Matrigel (Corning, Tewksbury, MA). 24 animals were implanted with unilateral flank tumors, and 6 animals were implanted with bilateral flank tumors. The animals underwent treatment 20 days after implantation.

Radiotracer Preparation

[0162] The preparation of ⁸⁹Zr-labeled liposomes is described by Perez-Medina et al in J Nucl Med.2014 Oct;55(10):1706-11. Briefly, pegylated liposomes containing the phospholipid chelator DSPE-DFO were prepared by the sonication method. The mean diameter of the liposomes was 105.8 ± 5.5 nm, whereas the mean diameter of Doxil was 82.4 ± 0.2. For radiolabeling, a solution of 0.3 % DFO-bearing liposomes in PBS was reacted with ⁸⁹Zr-oxalate at 40 °C for 2 h. The labeled liposomes were separated from free, unreacted ⁸⁹Zr through spin filtration.

Experimental Design and Treatment

[0163] The experiments were performed following guidelines in an Institutional Animal Care and Use Committee approved protocol. Details of animal numbers, cohorts, sacrifice time point, and measurements performed in each cohort are provided in Table 2.

[0164] The uptake behavior of exemplary tracer ⁸⁹Zr-NRep was used to quantify the effect of electric pulses on the permeability and retention properties of tumors in mice bearing unilateral tumors (n=24). Simultaneously, the uptake of the ⁸⁹Zr-NRep was compared with uptake of liposomal doxorubicin between treated and untreated tumors. Mice bearing bilateral tumors (n=6) were used to quantify the cytotoxic and electroporation mediated effects of treatment with electric pulses. These mice also underwent imaging (PET and autoradiography) to visualize the distribution of the ⁸⁹Zr-NRep within the tumors. All treatments were delivered using two 21 G stainless steel needles placed into the tumors. The needles were placed parallel to each other at a distance of 5-7mm contingent on the size of the tumor. The length of the needle was insulated except for 5mm at the tip that was left uncovered to allow passage of electricity. Tumors were treated with pulses with sufficient voltage to induce an electric field with strength of 700V/cm between the needle electrodes. Each animal was treated with 10 pulses delivered at 1 Hz with a pulse length of 90 microseconds. ECM835 generator (BTX, Harvard Apparatus, Holliston, MA) was used to deliver the square wave pulses. The pulse parameters were chosen for their ability to induce reversible electroporation of the tumor with limited cytotoxic effects. Animals with unilateral tumors received an injection of the ⁸⁹Zr-NRep (28.4 ± 0.4 μCi) and liposomal doxorubicin (0.2 mg) one hour post-treatment. Animals with bilateral tumors received injection of just the ⁸⁹Zr-NRep, either one hour before (n=3) or after (n=3) treatment. All animals were recovered post-treatment and were kept under regular observation until designated sacrifice time points.

Measurements and Imaging

[0165] Animals were sacrificed at time points as detailed in Table 2. Post-mortem, tumor samples from mice bearing unilateral tumors were carefully extracted and weighed. Tumors were then either divided into 2-3 smaller pieces or kept intact depending on the relative size of the tumor. The radiation in these tumor samples was counted using a Wizard 2480 Automatic Gamma Counter (Perkin Elmer, Waltham, MA). Quantification of doxorubicin in the samples was performed as previously reported (See Laginha KM, et al., Clin Cancer Res.2005 Oct 1;11(19 Pt 1):6944-9). Immediately after gamma counting, tumor samples were

homogenized in lysis buffer (10:1 v/w ratio) and were processed for drug extraction. Samples (200 µL) were added to a 96 well plate and drug measurements were performed using a microplate reader (Safire, Tecan, Mannedorf, Switzerland). A calibration curve was generated by adding increasing known quantities of doxorubicin to tumor sample lysates from untreated animals (no drug or pulse delivery). Animals with bilateral tumors underwent PET imaging at 2 hours and 24 hours following injection of ⁸⁹Zr-NRep. Imaging was performed using an Inveon MicroPET/CT (Siemens Healthcare Global). Whole body PET static scans recording a minimum of 50 million coincident events were performed, with duration of 10-20 min. The image data was normalized to correct for nonuniformity of response, dead-time count losses, positron branching ratio, and physical decay to the time of injection, but attenuation, scatter, or partial-volume averaging correction was not performed. The counting rates in the PET images were converted to equivalent activity concentration (percentage injected dose per gram of tissue) through use of a system calibration factor. Images were analyzed using ASIPro VMTM software (Concorde Microsystems). Quantification of activity concentration was done by averaging the maximum values in at least 5 ROIs drawn on adjacent slices of the treated and untreated tumors. Tumors were excised and embedded in OCT (Sakura Finetek, Torrance, CA), frozen and sectioned in 10 µm thick sections at five different levels throughout the tumor. Sections were imaged against a phosphor imaging plate (BASMS-2325, Fujifilm, Valhalla, NY) and the plates were read on a Typhoon 7000IP plate reader (GE Healthcare, Pittsburgh, PA) at a pixel resolution of 25 µm.

Histology

[0166] Tissue sections from animals bearing bilateral tumors (treated=6, control=6) underwent histology analysis. OCT embedded and sectioned frozen tumor samples were then stained with IBA1, Cleaved Caspase-3, and Hematoxilin and Eosin. An experienced staff veterinary pathologist evaluated the samples for evidence of necrosis or injury, and to ascertain differences in macrophage population between treated and untreated tumor samples.

Statistics

[0167] Data was compiled and descriptive statistical measures such as the mean and standard deviation was calculated. In animals with unilateral tumors, uptake of tracer ⁸⁹Zr-NRep and drug liposomal doxorubicin between treated and untreated tumors was compared using the non-parametric Wilcoxon rank sum test at each timepoint (6, 24 and 48 hours post-treatment). The uptake behavior of tracer ⁸⁹Zr-NRep and liposomal doxorubicin was correlated by fitting a linear regression line. Data handling and statistical analysis was automated using software wherever possible (Matlab, Mathworks Natick MA).

Results

Drug and ⁸⁹Zr-NRep Uptake

[0168] When compared to untreated control tumors, treated tumors demonstrated significantly increased tracer / drug uptake at the 6 hour time point. The changes were not statistically significant at the 24-hour and 48 hour time points, suggesting that the post-treatment increase in uptake did not persist at later time points (FIG.15A-F and Table 3). Treatment accelerated the uptake of exemplary tracer ⁸⁹Zr-NRep (FIG.16A). Measurements performed at the 6 hour time point indicated that treated tumors had 78.8±24% of their maximum ⁸⁹Zr-NRep uptake seen at 48 hours. In comparison, untreated tumors had just 41±15.6% of the maximum uptake seen at 48 hours. Similar trends were observed in drug uptake (FIG.16B), where measurements performed at the 6 hour time point indicated that treated tumors had 89.1±34.9% of their maximum drug uptake seen at 48 hours. In comparison, untreated tumors at the 6 hour time point had just 30.8±26.8% of the maximum uptake seen at 48 hours. Both the tracer and drug exhibited a linear increase in uptake with time (FIG.16C & D) in all treated tumors. There was no statistical difference between tracer and drug uptake at all time points. Evaluation indicated a high degree of correlation in both treated (r²=0.84) and untreated tumors (r²=0.8). Uptake of drug in tumors had limited correlation with the size of the tumor at all three assessed time points in both treated (FIG.17 A-C) and untreated tumors (FIG.17 D-F)

[0169] Electroporation Mediated Drug Delivery Can Be Monitored with a Radiotracer Uptake of exemplary tracer ⁸⁹Zr-NRep in treated and untreated tumors appeared similar to that of drug (e.g., doxorubicin), with significantly greater uptake at 6 and 24 hours timepoints in treated tumors, but no significant difference in overall uptake at 48 hours following injection (FIG.15). Similar to drug, electroporation accelerated uptake of tracer in treated tumors. Measurements performed at the 6 hour time point indicated that treated tumors had 78.8 ± 24% of their maximum tracer uptake (complete elimination from blood pool at 48 hours). In comparison, untreated tumors had just 41 ± 15.6% of the maximum uptake seen at 48 hours (FIG.16A). Both tracer and drug exhibited a linear increase in uptake with time (FIG.16 C & D) in both treated and untreated tumors. There was no statistical difference between tracer and drug uptake at all time points. Evaluation indicated a high degree of correlation drug and tracer in both treated (r² = 0.84) and untreated tumors (r² = 0.8). Uptake of tracer also had limited correlation with the tumor weight at all three time points in both treated (FIG.17 A-C) and untreated tumors (FIG.17 D-F).

PET Imaging and Autoradiography

[0170] Presence of exemplary tracer ⁸⁹Zr-NRep in the blood pool during pulse delivery resulted in immediate uptake of nanoparticles on PET imaging (FIG.18 A, solid arrow; 10.57 ± 0.95 %ID/g). Uptake was limited in contralateral untreated tumors (FIG.18 A and B, dashed arrow; 3.59 ± 0.45 %ID/g) and in tumors where ⁸⁹Zr-NRep was injected one hour after pulse delivery (FIG.18 B, solid arrow; 6.61 ± 0.85 %ID/g). However, PET imaging 24 hours following injection suggested uptake to be similar in tumors undergoing electroporation treatment after or before injection of tracer ⁸⁹Zr-NRep (FIG.18 C and D, solid arrows), while contralateral untreated tumors (FIG.18 C and D, dashed arrows) demonstrated markedly lesser concentration of ⁸⁹Zr-NRep (Inject/RE: 13.16 ± 0.69 %ID/g vs. RE/Inject: 12.84 ± 7.18 %ID/g vs. Contralateral Control: 3.9 ± 1.3 %ID/g; p < 0.01). Overall, when compared to untreated contralateral tumors electroporation, treatment increased the deposition of exemplary tracer ⁸⁹Zr- NRep in treated tumors, independent of the relative timing of tracer injection and pulse delivery. PET imaging suggested that treatment resulted in uniform distribution (FIGs.18 and 19A) of the ⁸⁹Zr-NRep within the tumor. These findings were confirmed with post-mortem

autoradiography imaging of tumors (FIG.19 B and C, and FIG.20 A) and comparing radiation counts in treated tumors with contralateral control (FIG.20 B).

Histology

[0171] Pathologist interpretation of H&E stains suggested tissue injury and cell death in the immediate vicinity of needle placement (FIG.21). However, this zone of injury was small and restricted to within 1 mm of the location of needle placement. Comparison of cleaved Caspase-3 stains between treated and contralateral control tumors suggested focal regions of increased staining adjacent to location of needle placement, consistent with observations from H&E stains. Review of IBA1 stained samples did not suggest increased presence of

macrophages in treated tumors when compared with the untreated tumor from the contralateral side.

Discussion

[0172] Without wishing to be bound by theory, the results from the experiments appear to demonstrate that in addition to electroporation of cells, electric pulses can modify the tumor microenvironment. Without wishing to be bound by theory, the passive or secondary effects of electric pulse delivery seems to alter the permeability of microvasculature (See Sersa G, et al., Eur J Cancer.1999 Apr;35(4):672-7.; Markelc B, et al., J Membr Biol.2012 Sep;245(9):545-54; Bellard E, et al. J Control Release.2012 Nov 10;163(3):396-403.), and the fluid redistribution may change diffusion characteristics within the tumor itself (See Zhang Zi Li W et al.,

Nanomedicine (Lond).2014;9(8):1181-92). Without wishing to be bound by theory, the results of this study demonstrate that electroporation related changes in vascular permeability may be another important mode for delivery of nanoparticles into tumors. Without wishing to be bound by theory, cumulatively, these changes were seen to affect the extravasation and accumulation behavior of nanoparticle agents. The effects of electric pulses were mapped with the novel liposomal radiotracer, and it was found that liposomal doxorubicin injected intravenously demonstrated uptake behavior identical to ⁸⁹Zr-NRep in both treated and untreated tumors.

While treatment with electric pulses did not increase the overall uptake of tracer ⁸⁹Zr-NRep or drug within the tumor, the net effect of treatment was observed to reduce the time required to reach maximum uptake values. Without wishing to be bound by theory, mice receiving treatment with electric pulses freely available nanoparticles (⁸⁹Zr-NRep or Drug) may be clearing from the blood pool earlier than in untreated mice, which may explain the absence of increased uptake in treated mice at 24 and 48 hour time points. Further, autoradiography analysis and PET imaging performed on a subset of animals indicated that distribution of the tracer was more uniform and widespread than in untreated tumors. The tumor microenvironment can be heterogeneous, demonstrating non-uniform deposition of nanoparticle agents in different regions. Without wishing to be bound by theory, in certain embodiments, application of electric pulses may normalize the microenvironment by creating a baseline level of permeability and retention in regions experiencing the effect of the electric pulses.

[0173] Comparison of the relative order of treatment and tracer injection suggested that injection of tracer before pulse delivery elicited an immediate (2 hours post-treatment) increase in tracer uptake. Such behavior was not observed where treatment preceded tracer injection by one hour, which was the main experimental protocol. Without wishing to be bound by theory, the gap between pulse delivery and tracer injection allowed sufficient time to complete resealing of all electroporation mediated membrane pores. However, measurements taken at the 24 hour time point demonstrated no differences in uptake between tumors from either group (injection before treatment or injection following treatment). Without wishing to be bound by theory, this suggests that the majority of uptake may be driven by the passive effects of electric pulse treatment. In certain embodiments, using this technology, it may be possible to increase the dose of drug (e.g., Doxil) delivered to tumor tissue. Without wishing to be bound by theory, in certain embodiments, during electroporation, the cells are in a permeabilized state for a short time window (few minutes) within which drug delivery has to be completed and/or during which drug delivery is enhanced. In the proposed approach it was possible to decouple delivery of electric pulses from the delivery of the therapeutic agent, removing the constraint of completing treatment delivery within a short span of time.

[0174] This work demonstrates that secondary effects of electric pulses may also support delivery of nanoparticles without requiring acute timing to match the window when the cells are in a permeable state. Electric pulses based nanoparticle delivery provides added benefits of short treatment time (<1 minute), and the use of non-ionizing energy to improve permeability of tumor vasculature.

[0175] The results of the presented study also demonstrate that companion imaging agents, such as the tracer ⁸⁹Zr-NRep, can be used to monitor and predict electroporation mediated uptake of nanoparticle therapeutics in vivo. This enhances the translation potential of this combination approach for future studies in patients. Monitoring tissue effects of

electroporation, such as membrane permeabilization and vascular changes, are difficult to accurately monitor with common imaging techniques. Superparamagnetic and radiolabeled nanoparticles have been evaluated as contrast agents for imaging electroporation mediated nanoparticle delivery with MRI and PET imaging techniques. However, the clinical utility of these agents were limited as they were either integrated with the therapeutic or were not completely validated for their ability to act as a reporter for therapeutics used with patients.

[0176] The results presented here, however, demonstrate that companion imaging reporters can be used to monitor and predict drug delivery using reversible electroporation.

[0177] Comparing nanoparticle accumulation following electroporation in bilateral and unilateral tumors indicates that that the vascular effect of electroporation can accelerate nanoparticle deposition. Effectively, this can increase the concentration or dose of the therapeutic in a very short period of time. In the bilateral tumor model, faster tracer (⁸⁹Zr-NRep) deposition in electroporation treated tumors led to competitive uptake when compared to untreated contralateral tumors. In turn, this led to 2-3 fold increase in nanoparticle uptake in treated tumors when compared to untreated controls. In unilateral tumors, this manifested as accelerated uptake in electroporation treated tumors with maximum uptake being reached within six hours of treatment. In comparison, without wishing to be bound by theory, it is estimated that in mice with untreated tumors the long circulation time (~48 hours) and slow clearance of drug from the blood pool allowed uptake in these tumors to catch up with that of electroporation treated mice. From a therapeutic perspective, rapid accumulation of drug in electroporation treated tumors can increase the concentration of the drug within the tumor and may enhance the cytotoxic effect of such therapeutics. Also, electroporation may be suited for delivery of nanoparticles that clear rapidly (<24 hours) from the blood pool.

[0178] In conclusion, treatment with electric pulses opens a simple and rapid way of altering the tumor microenvironment for enhancing the delivery of nanoparticle therapeutics. Without wishing to be bound by theory, while in certain embodiments the technique does not substantially increase the overall uptake of nanoparticles, it seems that it alters the rate and dynamics of uptake. Ultimately, such a treatment could therefore help to reach drug tissue levels in a tumor necessary to achieve better treatment outcomes, and could become standard of treatment during interventional ablation procedures. EXAMPLE 3

[0179] In this study, the feasibility and acute safety following catheter directed endoluminal in vivo irreversible electroporation (IRE) of porcine bronchus was evaluated.

Background and Objective.

[0180] Thermal ablation of tumors adjacent to large airways presents risk of

bronchopleural fistula, stenosis or stricture formation. The heat sink effect of large airways can also impact treatment outcomes. IRE is unaffected by the heat sink effect and has good safety profile when used adjacent to hollow organs in patients. The purpose of this study was to evaluate feasibility and intra-procedural safety of our new catheter electrode device for endobronchial IRE.

Materials and Methods

[0181] An expandable catheter electrode was designed to allow circumferential contact with airways of any diameter (See FIG.22). Endobronchial pulse delivery (treatment settings) was performed in the left or right main bronchi at 9 locations in 7 swine. Catheter placement was performed under fluoroscopy guidance and post-treatment CT was performed in all animals (See FIG.23). Animals were sacrificed 4 hours after ablation; airway and surrounding parenchyma was extracted for immunohistochemistry. CT images were used to create numerical simulations to estimate treatment zone and thermal effects.

Results [0182] Treatment was successfully completed in all animals. Infiltrative opacity surrounding ablated bronchus was seen on post treatment CT. There was no sign of airway perforation (See FIG.23). Measurements on gross pathology indicated ablation of length 2.6±0.2cm with circumferential penetration of 2.2±0.1cm from the bronchial wall into the parenchyma (See FIG.24). Sloughing of bronchial epithelium, and interstitial edema and hemorrhage were observed in the parenchyma in histology specimens (See FIG.25). Clear ablation margins were difficult to establish because of the early status of cell death in specimens. Simulations suggest preferential passage of electric field into the parenchyma and temperature increase restricted to the immediate vicinity of the electrodes (<2 mm) (See FIG.26).

Conclusion

[0183] Catheter directed endobronchial IRE may provide an alternate to thermal ablation for treatment of tumors adjacent to large airways. As the endobronchial approach directs electrical energy into the tissue it may reduce distortive effects associated with IRE during percutaneous treatment delivery in the lung. Further examination should be done to confirm the exact ablated area or late onset complication.

Claims

What is claimed is: 1. A method of enhancing uptake of an administered composition into a tissue of interest, the method comprising:

administering to a subject a therapeutic and/or diagnostic agent; and

delivering electric energy (e.g., one or more electric pulses) to an interior surface of a body lumen of the subject (e.g., at one or more points/positions about a circumference of the lumen), thereby enhancing uptake of the administered therapeutic and/or diagnostic agent into the tissue of interest.

2. The method of claim 1 wherein the tissue of interest is within, on, and/or in the vicinity of the interior surface of the body lumen of the subject.

3. The method of claim 2, wherein the body lumen is the interior of a vessel (e.g., an artery, vein, gastrointestinal tract, esophagus, alimentary canal, respiratory tract, nasal passage, bronchi in the lungs, renal tubule, urinary collecting duct, female genital tract, pathway of the vagina, uterus, fallopian tube, the stomach, sinus tract, biliary duct, pancreatic duct, breast duct, and/or the abdominal cavity).

4. The method of any one of claims 1-3, wherein the tissue is a manifestation of neoplastic disease.

5. The method of any one of claims 1-4, wherein the neoplastic disease is cancer.

6. The method of any one of claims 1-5, wherein the tissue of interest is a dense tumor.

7. The method of any one of claims 1-6, wherein the therapeutic and/or diagnostic agent comprises a nanoparticle.

8. The method of any one of claims 1-7, wherein the therapeutic and/or diagnostic agent comprises a liposome.

9. The method of any one of claims 1-8, wherein the electric energy is delivered before administration of the therapeutic agent.

10. The method of any one of claims 1-9, wherein the electric energy is delivered after administration of the therapeutic agent.

11. The method of any one of claims 1-10, wherein the electric energy is delivered using a device of any one of claims 38-48.

12. The method of any one of claims 1-11, wherein the electric energy is delivered by one or more electrodes in the form of one or more electric pulses, wherein the electric pulses are wave pulses (e.g., square waves, sine waves, step waves, triangle waves, or sawtooth waveforms).

13. The method of claim 12, wherein the electric pulses are square wave pulses.

14. The method of claim 12 or 13, wherein the number of pulses applied at a given position along the lumen (e.g. at a given general location of the electrodes) is between 1 and 1000, between 5 and 500, between 10 and 100, or between 10 and 50.

15. The method of any one of claims 12-14, wherein the pulse frequency is between 0.1 Hz and 20 Hz (e.g., between any two of the following values: 0.1 Hz, 0.2 Hz, 0.5 Hz, 1 Hz, 2 Hz, 3 Hz, 4 Hz, 5 Hz, 10 Hz, and 20 Hz).

16. The method of any one of claims 12-15, wherein the pulse width is between 0.001 µs and 1 s, between 0.01 µs and 100 ms, between 0.1 µs and 10 ms, between 1 µs and 1 ms, or between

17. The method of any one of claims 12-16, wherein the pulses have sufficient voltage to induce an electric field between the electrodes, wherein the electric field has a strength of at least 100 V/cm, at least 200 V/cm, at least 300 V/cm, at least 400 V/cm, at least 500 V/cm, at least 600 V/cm, at least 700 V/cm, at least 800 V/cm, at least 900 V/cm, or at least 1000 V/cm.

18. The method of any one of claims 12-17, wherein the voltage applied is between 1 and 1000 V, between 5 and 500 V, between 10 and 100 V, or between 10 and 50 V.

19. The method of any one of claims 12-18, wherein the electrodes are needle electrodes.

20. The method of any one of claims 12-19, wherein the electrodes are in direct contact with tumor tissue.

21. The method as in any one of claims 12-18, comprising delivering the electric pulses using a device of any one of claims 38-48.

22. A method for treating a condition comprising delivering electric square wave pulses to tissue of a luminal organ (e.g., an artery, vein, gastrointestinal tract, esophagus, alimentary canal, respiratory tract, nasal passage, bronchi in the lungs, renal tubule, urinary collecting duct, female genital tract, pathway of the vagina, uterus, fallopian tube, the stomach, and the abdominal cavity) or other vessel of a subject in a circumferential or focal fashion.

23. The method of claim 22, wherein the square wave pulses ablate tumors or other undesirable tissue within the organ or other vessel.

24. The method of any one of claims 22-23, comprising performing irreversible

electroporation and/or nanoporation of the tissue.

25. The method of any one of claims 22-24, comprising performing reversible

electroporation of the tissue.

26. The method of any one of claims 22-25, wherein the tissue is an interior wall of a lumen.

27. The method of any one of claims 22-26, comprising transfecting the tissue with genetic material and/or one or more drugs.

28. The method of any one of claims 22-27, wherein the electric square wave pulses are delivered using a device of any one of claims 38-48.

29. A method for treating a condition comprising delivering high frequency electrical energy into the inner walls of a luminal organ.

30. The method of claim 29, comprising performing coagulation of bleeding in the luminal organ.

31. The method of any one of claims 29-30, wherein the condition is a varix (or varices), internal hemorrhage, or a bleeding ulcer.

32. The method of any one of claims 29-31, wherein the high frequency electrical energy is delivered using a device of any one of claims 38-48.

33. A method for treating a condition comprising delivering radiofrequency energy to a lumen wall of a hollow organ.

34. The method of claim 33, further comprising partial ablation of the lumen wall.

35. The method of any one of claims 33-34, wherein the condition is hypertrophy of smooth muscle or muscularis of luminal organs.

36. The method of any one of claims 33-35, wherein the condition is asthma, esophageal strictures, or vascular stenosis.

37. The method of any one of claims 33-36, wherein the radiofrequency energy is delivered using a device of any one of claims 38-48.

38. A device for treating and/or diagnosing a condition in a subject, the device comprising a catheter comprising an expandable element at a distal end that maintains contact with the interior surface of a body lumen at at least two points about a circumference of the body lumen, the expandable element capable of delivering electric energy (e.g., one or more electric pulses) at the at least two points.

39. The device of claim 38 wherein the expandable element has an adjustable diameter such that the expandable element is capable of changing diameter (increasing and/or decreasing diameter) as the catheter is drawn along a length of the body lumen, so as to maintain contact between the body lumen at at least two points as the catheter is drawn along the length of the body lumen].

40. The device of any one of claims 38-39, wherein the expandable element comprises an electrically conducting material.

41. The device of any one of claims 38-40, wherein the expandable element is disposed concentrically around substantially the (e.g., entire) outer circumference of the catheter.

42. The device of any one of claims 38-41, wherein the expandable element is disposed around a fraction of the circumference of the catheter.

43. The device of any one of claims 38-42, wherein the expandable element is a basket.

44. The device of any one of claims 38-43, further comprising a second expandable element.

45. The device of any one of claims 38-44 further comprising a handle, wherein the handle can be manipulated to cause the expandable element to expand from a first, compressed state to a second, expanded state.

46. The device of any one of claims 38-45, wherein the expandable element has a fully expanded circumference at its widest of at least approximately 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm, 8 cm, 9 cm, 10 cm, 11 cm, 12 cm, 13 cm, 14 cm, 15 cm, 16 cm, 17 cm, 18 cm, 19 cm, 20 cm, 22 cm, 24 cm, 26 cm, 28 cm, 30 cm or more.

47. The device of any one of claims 38-45, wherein the expandable element has a fully expanded circumference at its widest of between 1 cm and 100 cm, of between 3 cm and 75 cm, of between 5 cm and 50 cm, of between, 7 cm and 30 cm, or between 10 cm and 20 cm.

48. The device of any one of claims 38-47, wherein the expandable element has an adjustable diameter or an adjustable shape after expansion within the body lumen (e.g., in certain embodiments, the expandable element is capable of decreasing diameter by at least

approximately 2%, 5%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 99% of its maximum diameter while still maintaining contact with the interior surface of a lumen of varying diameter, e.g., as the element is drawn through the lumen).

49. A composition comprising a nanoparticle for use in a method for treating a neoplastic disease in a subject, wherein the method comprises

administering to a subject having a tissue of interest the nanoparticle; and

delivering one or more electric pulses to an interior surface of a body lumen of the subject, thereby enhancing uptake of the administered therapeutic and/or diagnostic agent into the tissue of interest.

50. A composition comprising a nanoparticle comprising a radiolabel for use in a method of in vivo diagnosis of a neoplastic disease in a subject, wherein the method comprises:

administering to a subject having a tissue of interest the nanoparticle; and

51. A composition of any one of claims 49-50, wherein the nanoparticle is a liposome

52. A composition of any one of claims 49-51, wherein the electric pulses are delivered before administration of the nanoparticle.

53. A composition of any one of claims 45-48, wherein the electric pulses are delivered after administration of the nanoparticle.