CA2795401A1 - Treatment of sarcoidosis using placental stem cells - Google Patents

Treatment of sarcoidosis using placental stem cells Download PDF

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Publication number
CA2795401A1
CA2795401A1 CA2795401A CA2795401A CA2795401A1 CA 2795401 A1 CA2795401 A1 CA 2795401A1 CA 2795401 A CA2795401 A CA 2795401A CA 2795401 A CA2795401 A CA 2795401A CA 2795401 A1 CA2795401 A1 CA 2795401A1
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sarcoidosis
cells
placental cells
stem cells
express
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James Edinger
Steven Fischkoff
Aleksander Francki
Vladimir Jankovic
Bitao Liang
Philippe Martin
Cynthia Ray
Xiaokui Zhang
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Clarity Acquisition II LLC
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Anthrogenesis Corp
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/14—Antitussive agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
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    • A—HUMAN NECESSITIES
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    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
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    • A61P3/02—Nutrients, e.g. vitamins, minerals
    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
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Abstract

Provided herein are compositions and methods of treating individuals having sarcoidosis or a sarcoidosis-related disease or disorder, using placental cells, e.g., the placental stem cells and placental multipotent cells (PDACs) described herein. Also provided herein are kits comprising said cells or a composition thereof.

Description

TREATMENT OF SARCOIDOSIS USING PLACENTAL STEM CELLS

1. FIELD

[0001] Provided herein are isolated placental cells (e.g., placental stem cells), populations of the placental stem cells, and methods of using the same to treat sarcoidosis or a sarcoidosis-related disease, disorder or condition, for example, systemic sarcoidosis, cutaneous sarcoidosis, Lofgren's syndrome, neurosarcoidosis, pulmonary sarcoidosis, cardiac sarcoidosis, ocular sarcoidosis, or sarcoidosis with the involvement of other organs or tissues.
2. BACKGROUND

[0002] Sarcoidosis, also called sarcoid disease or Besnier-Boeck disease, is a multisystem granulomatous inflammatory disease that can affect a variety of organs (e.g., lungs, skin, heart, eyes, liver, nervous system, kidneys, or musculoskeletal system). The etiology of sarcoidosis has remained unknown, though it has been linked to alterations in the immune response after exposure to an environmental, occupational, or infectious agent in genetically susceptible individuals.
[0003] Steroid therapy (e.g., corticosteroids such as prednisone) has been the standard treatment for sarcoidosis. However, the use of corticosteroids has a number of drawbacks.
For example, certain patients do not respond to steroid therapy. In addition, corticosteroids have several serious side effects, and their use is typically limited to progressive or severe conditions. Accordingly, there exists a need for new methods of treating sarcoidosis, using, for example, human placental stem cells.

3. SUMMARY
[0004] In one aspect, provided herein are methods of treating, managing, ameliorating or preventing sarcoidosis, or one or more sarcoidosis-related diseases, disorders and/or conditions, e.g., associated with, resulting in or caused by sarcoidosis. Also provided herein are kits for use in the treatment of sarcoidosis, or a disease, disorder or condition caused by, or relating to, sarcoidosis.
[0005] In one embodiment, provided herein is a method of treating an individual having sarcoidosis, or a sarcoidosis-related disease, disorder or condition, comprising administering to the individual a therapeutically effective amount of tissue culture plastic-adherent placental cells, e.g., placental multipotent cells or placental stem cells, also referred to herein as PDACs (placenta derived adherent cells, e.g., the placenta-derived adherent cells described in Section 4.2, below), or culture medium conditioned by PDACs (e.g., placental stem cells), in an amount and for a time sufficient for detectable improvement of one or more symptoms of sarcoidosis, or said disease, disorder or condition.
[0006] In another aspect, provided herein is the use of placental cells (e.g., placental stem cells) in the manufacture of a medicament for treating, managing, ameliorating or preventing sarcoidosis or diseases, disorders and/or conditions caused by, or relating to, sarcoidosis.
[0007] In another embodiment, provided herein are kits comprising PDACs or a therapeutic cell composition thereof, which can be prepared in a pharmaceutically acceptable form, for example by mixing with a pharmaceutically acceptable carrier, and an applicator, along with instructions for use.
[0008] In certain embodiments, the method comprises administering placental cells (e.g., placental stem cells) to said individual in an amount and for a time sufficient for detectable improvement of one or more symptoms of said sarcoidosis or said disease or disorder, as compared to the individual prior to administration of said placental stem cells.
[0009] In certain embodiments, said therapeutically effective amount of placental cells reduces, e.g., detectably reduces, the number of, or degree of severity of, or reduces the rate of increase in the number of, or degree of severity, said one or more symptoms in said individual, e.g., and/or detectable improvement, of one or more symptoms of sarcoidosis or sarcoidosis-related diseases, disorders or conditions. In specific embodiments, said therapeutically effective amount of placental cells reduces, e.g., detectably reduces, the number of, or degree of severity of, or reduces the rate of increase in the number of, or degree of severity, one or more granulomas in said individual. In one embodiment, said therapeutically effective amount is an amount that results in a detectable improvement in at least one symptom of said sarcoidosis.
[0010] In one specific embodiment, said sarcoidosis is systemic sarcoidosis, and said symptoms comprise one ore more of formation of granulomas, fatigue, weight loss, fever, aches, pains, arthritis, dry eyes, swelling of the knees, blurry vision, shortness of breath, cough or skin lesions.
[0011] In another specific embodiment, said sarcoidosis is cutaneous sarcoidosis, and said symptoms comprise a skin lesion. In a more specific embodiment, said cutaneous sarcoidosis comprises one or more of annular sarcoidosis, erythrodermic sarcoidosis, ichthyosiform sarcoidosis, hypopigmented sarcoidosis, morpheaform sarcoidosis, mucosal sarcoidosis, papular sarcoid, scar sarcoid, subcutaneous sarcoidosis or ulcerative sarcoidosis. In a more specific embodiment, said skin lesion comprises one or more of erythema nodosum, nummular eczema, erythema multiforme, calcinosis cutis and pruritus, lupus pernio, skin plaques, maculopapular eruptions, nodular lesions deeper in the skin or infiltration of old scars.
[0012] In another specific embodiment, said sarcoidosis is Lofgren's syndrome, and said symptoms comprise one or more of erythema nodosum, bilateral hilar denopathy, arthritis, arthralgias or fever. In another specfic embodiment, said sarcoidosis is neurosarcoidosis, and said symptoms comprise one or more of formation of granulomas in the nervous system, headache, confusion, malaise or facial paralysis. In another specific embodiment, said sarcoidosis is pulmonary sarcoidosis, and said symptoms comprise one or more of cough, shortness of breath, chest pain, a feeling of tightness in the chest, hoarseness, nasal obstruction and recurrent or persistent sinusitis. In another specific embodiment, said sarcoidosis is cardiac sarcoidosis, and said symptoms comprise one or more of chest pain, palpitations, congestive heart failure, pericarditis or papillary muscle dysfunction, shortness of breath, ankle swelling, irregular heart beat or sudden death. In another specific embodiment, said sarcoidosis is ocular sarcoidosis, and said symptoms comprise one or more of red or watery eyes, granulomatous uvetis, iris nodules, retinochoroiditis, conjunctivitis, lacrimal gland involvement or proptosis. In another specific embodiment, said sarcoidosis is sarcoidosis with the involvement of other organs or tissues, e.g., sarcoidosis with musculoskeletal, hepatic, hematologic, psychiatric, renal, splenic, nasal sinus, oral, gastric or intestinal, endocrine, pleural or reproductive involvement. In another specific embodiment, said sarcoidosis is sarcoidosis of one or more organs or tissues other than pulmonary tissues or organs. In a more specific embodiment, said sarcoidosis is sarcoidosis of one or more organs or tissues other than the lungs.
[0013] In certain embodiments, the method of treatment comprises administering at least about 1 x 107, 5 x 107, 1 x 108, 5 x 108, 1 x 109, 5 x 109, or 1 x 1010 placental cells to said individual. In another specific embodiment, said placental cells have been proliferated in vitro for no more than 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 population doublings. In another embodiment of any of the embodiments herein, said placental cells have been cryopreserved and thawed prior to said administering.
[0014] In certain embodiments of any of the above methods, said placental cells are adherent to tissue culture plastic and are CD34-, CD10+, CD105+ and CD200+ as detected by flow cytometry. In a specific embodiment, the placental cells have the capacity to differentiate into osteogenic or chondrogenic cells. In another embodiment, said placental cells are adherent to tissue culture plastic; CD34-, CD10+, CD105+ and CD200+ as detected by flow cytometry; and have the capacity to differentiate into cells having one or more characteristics of osteogenic or chondrogenic cells, e.g., characteristics of osteocytes or chondrocytes. In other embodiments, the placental cells additionally have the ability to differentiate into cells having one or more characteristics of neural cells or neurogenic cells, e.g., characteristics of neurons; one or more characteristics of glial cells, e.g., characteristics of glia or astrocytes;
one or more characteristics of adipocytic cells, e.g., characteristics of adipocytes; one or more characteristics of pancreatic cells; and/or one or more characteristics of cardiac cells.
[0015] In another embodiment, said placental cells are CD34-, CD10+, CD105+
and CD200+, and one or more of CD38-, CD45-, CD80-, CD86-, CD133-, HLA-DR,DP,DQ-, SSEA3-, SSEA4-, CD29+, CD44+, CD73+, CD90+, CD105+, HLA-A,B,C+, PDL1+, ABC-p+, and/or OCT-4+, as detected by flow cytometry and/or RT-PCR. In another embodiment, said placental cells are CD34-, CD45-, CD10+, CD90+, CD105+ and CD200+, as detected by flow cytometry. In another embodiment, said placental cells are CD34-, CD45-, CD10+, CD80-, CD86-, CD90+, CD105+ and CD200+, as detected by flow cytometry. In another embodiment, said placental cells are CD34-, CD45-, CD10+, CD80-, CD86-, CD90+, CD105+
and CD200+, and additionally one or more of CD29+, CD38-, CD44+, CD54+, SH3+
or SH4+, as detected by flow cytometry. In another embodiment, said placental cells are CD34-, CD38-, CD45-, CD10+, CD29+, CD44+, CD54+, CD73+, CD80-, CD86-, CD90+, CD105+, and CD200+ as detected by flow cytometry.
[0016] In another embodiment, said CD34-, CD10+, CD105+ and CD200+ placental cells are additionally one or more of CD11T, CD133-, KDR (VEGFR2-), HLA-A,B,C+, HLA-DP,DQ,DR, or Programmed Death-1 Ligand (PDL1)+, or any combination thereof. In another specific embodiment, said placental cells are CD34-, CD38-, CD45-, CD
10-'-, CD29-'-, CD44+, CD54+, CD73+, CD80-, CD86-, CD90+, CD105+, CD11T, CD133-, CD200+, KDR
(VEGFR2-), HLA-A,B,C+, HLA-DP,DQ,DR, or Programmed Death-1 Ligand (PDL I)-,-, as detected by flow cytometry.
[0017] In another embodiment, any of the placental cells described herein are additionally ABC-p+, as detected by flow cytometry, or OCT-4+ (POU5F1), e.g., as determined by RT-PCR, wherein ABC-p is a placenta-specific ABC transporter protein (also known as breast cancer resistance protein (BCRP) and as mitoxantrone resistance protein (MXR)). In another embodiment, any of the placental cells described herein are additionally SSEA3-or SSEA4-, e.g., as determined by flow cytometry, wherein SSEA3 is Stage Specific Embryonic Antigen 3, and SSEA4 is Stage Specific Embryonic Antigen 4. In another embodiment, any of the placental cells described herein are additionally SSEA3- and SSEA4-.
[0018] In another embodiment of the methods described herein, any of the placental cells described herein are additionally one or more of MHC-I+ (e.g., HLA-A,B,C+), MHC-II- (e.g., HLA-DP,DQ,DR) or HLA-G-. In another embodiment, any of the placental cells described herein are additionally each of MHC-I+ (e.g., HLA-A,B,C+), MHC-II- (e.g., HLA-DP,DQ,DR) and HLA-G-, as detected by flow cytometry.
[0019] In another embodiment, the CD34-, CD10+, CD105+, CD200+ cells are additionally one or more of CD29+, CD38-, CD44+, CD54+, CD80-, CD86-, SH3+ or SH4+. In another embodiment, the cells are additionally CD44+. In another embodiment, the CD34-, CD 10-'-, CD105+, CD200+ placental cells are additionally one or more of CD13+, CD29+, CD33+, CD38-, CD44+, CD45-, CD54+, CD62E-, CD62L-, CD62P-, SH3+ (CD73+), SH4+
(CD73+), CD80-, CD86-, CD90+, SH2+ (CD105+), CD106/VCAM+, CD11T, CD144/VE-cadherinb0v, CD184/CXCR4-, CD133-, OCT-4+, SSEA3-, SSEA4-, ABC-p+, KDR (VEGFR2-), HLA-A,B,C+, HLA-DP,DQ,DR, HLA-G-, or Programmed Death-1 Ligand (PDL I)-,-, or any combination thereof. In another embodiment, the CD34-, CD10+, CD105+, CD200+
placental cells are additionally CD13+, CD29+, CD33+, CD38-, CD44+, CD45-, CD54/ICAM+, CD62E-, CD62L-, CD62P-, SH3+ (CD73+), SH4+ (CD73+), CD80-, CD86-, CD90+, SH2+ (CD 105+), CD 106/VCAM+, CD 11 T, CD 144/VE-cadherind'm, CD

, CD133-, OCT-4+, SSEA3-, SSEA4-, ABC-p+, KDR (VEGFR2-), HLA-A,B,C+, HLA-DP,DQ,DR, HLA-G-, and Programmed Death-1 Ligand (PDL1)+, as detected by flow cytometry.
[0020] In other embodiments of the methods disclosed herein, the isolated placental cells are CD200+ and HLA-G-; CD73+, CD 105+, and CD200+; CD200+ and OCT-4+; CD73+, CD
105+
and HLA-G-; CD73+ and CD105+; or OCT-4+; or any combination thereof, as detected by flow cytometry.
[0021] In certain embodiments of the methods disclosed herein, the isolated placental cells are one or more of CD10+, CD29+, CD34-, CD38-, CD44+, CD45-, CD54+, CD90+, SH2+, SH3+, SH4+, SSEA3-, SSEA4-, OCT-4+, MHC-I+ or ABC-p+, where ABC-p is a placenta-specific ABC transporter protein (also known as breast cancer resistance protein (BCRP) and as mitoxantrone resistance protein (MXR)). In another embodiment, the isolated placental cells are CD10+, CD29+, CD34-, CD38-, CD44+, CD45-, CD54+, CD90+, SH2+, SH3+, SH4+, SSEA3-, SSEA4-, and OCT-4+. In another embodiment, the isolated placental cells are CD10+, CD29+, CD34-, CD38-, CD45-, CD54+, SH2+, SH3+, and SH4+. In another embodiment, the isolated placental cells are CD10+, CD29+, CD34-, CD38-, CD45-, CD54+, SH2+, SH3+, SH4+ and OCT-4+. In another embodiment, the isolated placental cells are CD10+, CD29+, CD34-, CD38-, CD44+, CD45-, CD54+, CD90+, MHC-1+, SH2+, SH3+, SH4+. In another embodiment, the isolated placental cells are OCT-4+ and ABC-p+. In another embodiment, the isolated placental cells are SH2+, SH3+, SH4+ and OCT-4+. In another embodiment, the isolated placental cells are OCT-4+, CD34-, SSEA3-, and SSEA4-.
In a specific embodiment, said OCT-4+, CD34-, SSEA3-, and SSEA4- cells are additionally CD 10+, CD29+, CD34-, CD44+, CD45-, CD54+, CD90+, SH2+, SH3+, and SH4+. In another embodiment, the isolated placental cells are OCT-4+ and CD34-, and either SH3+
or SH4+. In another embodiment, the isolated placental cells are CD34- and either CD10+, CD29+, CD44+, CD54+, CD90+, or OCT-4+. In certain embodiments, the isolated placental cells are CD10+, CD34-, CD105+ and CD200+.
[0022] In another embodiment, the isolated placental cells useful in the methods described herein are one or more of CD10+, CD29-, CD44+, CD45-, CD54/ICAM-, CD62-E-, CD62-L-, CD62-P-, CD80-, CD86-, CD103-, CD104-, CD105+, CD106/VCAM+, CD144/VE-cadherindim, CD 184/CXCR4-, (32-microglobulindim, MHC-Id'm, MHC-II-, HLA-Gdim, and/or PDLldim. In certain embodiments, such placental cells are at least CD29- and CD54-. In another embodiment, such isolated placental cells are at least CD44+ and CD106+. In another embodiment, such isolated placental cells are at least CD29+.
[0023] In certain embodiments of any of the above characteristics, expression of the cellular marker (e.g., cluster of differentiation or immunogenic marker) is determined by flow cytometry. In certain other embodiments, expression of the cellular marker is determined by RT-PCR.
[0024] In another embodiment, said placental cells useful in the methods disclosed herein, e.g., said CD10+, CD34-, CD 105-% CD200+ cells, express one or more genes at a detestably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes are one or more of ACTG2, ADARB1, AMIGO2, ARTS-l, B4GALT6, BCHE, Cl lorf9, CD200, COL4A1, COL4A2, CPA4, DMD, DSC3, DSG2, ELOVL2, F2RL1, FLJ10781, GATA6, GPR126, GPRC5B, ICAM1, IER3, IGFBP7, ILIA, IL6, IL18, KRT18, KRT8, LIPG, LRAP, MATN2, MEST, NFE2L3, NUAK1, PCDH7, PDLIM3, PKP2, RTN1, SERPINB9, ST3GAL6, ST6GALNAC5, SLC12A8, TCF21, TGFB2, VTN, and ZC3H12A, and wherein said bone marrow-derived mesenchymal stem cells have undergone a number of passages in culture equivalent to the number of passages said isolated placental cells have undergone. In certain embodiments, said expression of said one ore more genes is determined, e.g., by RT-PCR or microarray analysis, e.g, using a U133-A microarray (Affymetrix). In another embodiment, said isolated placental cells express said one or more genes when cultured for, e.g., anywhere from about 3 to about 35 population doublings, in a medium comprising 60% DMEM-LG (e.g., from Gibco) and 40%
MCDB-201 (e.g., from Sigma); 2% fetal calf serum (e.g., from Hyclone Labs.);
lx insulin-transferrin-selenium (ITS); lx linoleic acid-bovine serum albumin (LA-BSA); 10-dexamethasone (e.g., from Sigma); 10-4 M ascorbic acid 2-phosphate (e.g., from Sigma);
epidermal growth factor 10 ng/mL (e.g., from R&D Systems); and platelet-derived growth factor (PDGF-BB) 10 ng/mL (e.g., from R&D Systems). In another embodiment, said isolated placental cells express said one or more genes when cultured for from about 3 to about 35 population doublings in a medium comprising 60% DMEM-LG (e.g., from Gibco) and 40% MCDB-201 (e.g., from Sigma); 2% fetal calf serum (e.g., from Hyclone Labs.); lx insulin-transferrin-selenium (ITS); lx linoleic acid-bovine serum albumin (LA-BSA); 10-9 M
dexamethasone (e.g., from Sigma); 10-4 M ascorbic acid 2-phosphate (Sigma);
epidermal growth factor 10 ng/mL (e.g., from R&D Systems); and platelet-derived growth factor (PDGF-BB) 10 ng/mL (e.g., from R&D Systems).
[0025] In some embodiments, the placental cells express CD200 and do not express HLA-G, or express CD73, CD 105, and CD200, or express CD200 and OCT-4, or express CD73 and CD 105 and do not express HLA-G, or express CD73 and CD 105 and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising said stem cell when said population is cultured under conditions that allow for the formation of an embryoid-like body, or express OCT-4 and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising said stem cell when said population is cultured under conditions that allow for the formation of an embryoid-like body.
[0026] In certain embodiments, the placental cells express CD200 and ARTS1 (aminopeptidase regulator of type 1 tumor necrosis factor); ARTS-1 and LRAP
(leukocyte-derived arginine aminopeptidase); IL6 (interleukin-6) and TGFB2 (transforming growth factor, beta 2); IL6 and KRT 18 (keratin 18); IER3 (immediate early response 3), MEST
(mesoderm specific transcript homolog) and TGFB2; CD200 and IER3; CD200 and IL6;
CD200 and KRT18; CD200 and LRAP; CD200 and MEST; CD200 and NFE2L3 (nuclear factor (erythroid-derived 2)-like 3); or CD200 and TGFB2 at a detestably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells (BM-MSCs) wherein said bone marrow-derived mesenchymal stem cells have undergone a number of passages in culture equivalent to the number of passages said isolated placental cells have undergone. In other embodiments, the placental cells express ARTS-l, CD200, IL6 and LRAP;
ARTS-l, IL6, TGFB2, IER3, KRT 18 and MEST; CD200, IER3, IL6, KRT 18, LRAP, MEST, NFE2L3, and TGFB2; ARTS-1, CD200, IER3, IL6, KRT18, LRAP, MEST, NFE2L3, and TGFB2; or IER3, MEST and TGFB2 at a detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells BM-MSCs, wherein said bone marrow-derived mesenchymal stem cells have undergone a number of passages in culture equivalent to the number of passages said isolated placental cells have undergone.
[0027] In various embodiments, said isolated placental cells useful in the methods disclosed herein, e.g., placental stem cells or placental multipotent cells, are contained within a population of cells, at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the cells of which are said isolated placental cells. In certain other embodiments, the placental cells in said population of cells are substantially free of cells having a maternal genotype; e.g., at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the placental cells in said population have a fetal genotype, i.e., are fetal in origin. In certain other embodiments, the population of cells comprising said placental cells are substantially free of cells having a maternal genotype; e.g., at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the cells in said population have a fetal genotype, i.e., are fetal in origin. In certain other embodiments, the population of cells comprising said placental cells comprise cells having a maternal genotype; e.g., at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the cells in said population have a maternal genotype, i.e., are maternal in origin.
[0028] In an embodiment of any of the embodiments of placental cells (e.g., PDACs) herein, the placental cells facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising said isolated placental cells when said population is cultured under conditions that allow the formation of an embryoid-like body e.g., culture under proliferation conditions).
[0029] In certain embodiments of any of the placental cells disclosed herein, the cells are human.
[0030] In certain embodiments, any of the placental cells, e.g., placental stem cells or placental multipotent cells described herein, are autologous to a recipient.
In certain other embodiments, any of the placental cells, e.g., placental stem cells or placental multipotent cells described herein, are heterologous to a recipient.
[0031] In certain embodiments, the placental cells are cryopreserved prior to said administering. In another embodiment, said isolated placental cells are obtained from a placental cell bank (e.g., a PDAC bank).
[0032] In any of the above embodiments of isolated placental cells, the isolated placental cells generally do not differentiate during culturing in growth medium, i.e., medium formulated to promote proliferation, e.g., during proliferation in growth medium. In another embodiment, said isolated placental cells do not require a feeder layer in order to proliferate.
In another embodiment, said isolated placental cells do not differentiate in culture solely as the result of culture in the absence of a feeder cell layer.
[0033] In certain embodiments, said isolated placental cells are obtained by perfusion of a post-partum placenta that has been drained of blood and perfused to remove residual blood;
drained of blood but not perfused to remove residual blood; or neither drained of blood nor perfused to remove residual blood. In another specific embodiment, said isolated placental cells are obtained by physical and/or enzymatic disruption of placental tissue.
[0034] Cell surface, molecular and genetic markers of placental cells, useful in the methods provided herein, are described in detail in Section 4.2, below.
[0035] In specific embodiments, said individual to whom placental stem cells are administered is additionally administered one or more of a second therapy (e.g., second therapeutic agent), wherein said second therapy (e.g., second therapeutic agent) comprises anti-inflammatory agents, steroids, immune suppressants, antibiotics, TNF-a inhibitors (e.g., infliximab, adalimumab), PDE-4 inhibitors (e.g., apremiliast), IL-12 inhibitors, IL-23 inhibitors, aviptadil, IFN-y inhibitors, or antimalarials (e.g., chloroquine or hydroxychloroquine).
[0036] Examples of anti-inflammatory drugs useful in the treatment of sarcoidosis, or sarcoidosis-related diseases, disorders or conditions include, but are not limited to, mesalamine, 5-ASA (5-aminosalicylic acid) agents (e.g., ASACOL (mesalamine, delayed-release), DIPENTUM (Osalazine), PENTASA (mesalamine controlled-release)), sulfasalazine (a combination of 5-ASA and sulfapyridine), anti-inflammatory antibodies (e.g., Infliximab (REMICADE )), and the like. Examples of steroids useful in the treatment of sarcoidosis, or sarcoidosis-related diseases, disorders or conditions include, but are not limited to, cortisone, hydrocortisone, predisone, methylprednisone, and the like. Typically, as practiced in the art, the dosage of steroid is first delivered in a relatively large dose, followed by smaller dosages as inflammation subsides. Examples of immune suppressants useful in the treatment of sarcoidosis, or sarcoidosis-related diseases or disorders include, but are not limited to, cyclosporine A, 6-mercaptopurine or azathioprine. Any antibiotic can be used in the treatment of Crohn's disease, including, e.g., ampicillin, sulfonamide, cephalosporin, tetracycline, and/or metronidazole. In another specific embodiment, the second therapy is an administration of porcine whipworms, e.g., ova of Trichuris suis.
[0037] In another embodiment, administration of placental cells (e.g., PDACs), or therapeutic compositions comprising such cells, to an individual in need thereof, can be accomplished, for local or systemic administration, by transplantation, implantation (e.g., of the cells themselves or the cells as part of a matrix-cell combination), injection (e.g., directly to the site of the disease or condition, for example, directly to an granuloma), infusion, delivery via catheter, or any other means known in the art for providing cell therapy. In other embodiment, placental cells (e.g., PDACs), or therapeutic compositions comprising such cells can be delivered to an individual by intravenous, intraarterial, intraperitoneal, intraventricular, intraurethral, intraaternal, intracranial, intramuscular, intrasynovial, intraocular, intravitreal (e.g., where there is an ocular involvement), intracerebral, intracerebroventricular (e.g., where there is a neurologic or brain invovlement), intracardiac (e.g., where there is a cardiac involvement), intrathecal,intraosseous infusion, intravesical, and transdermal, intracisternal, epidural, subcutaneous administration.
[0038] In specific embodiments, the placental cells are administered locally, e.g., directly into, or adjacent to (e.g., within 1-5 cm of) one or more damaged tissues caused by, or relating to, sarcoidosis. In specific embodiments, the placental cells are administered intralesionally, e.g., directly into, or adjacent to (e.g., within 1-5 cm of) one or more lesions or granulomas caused by, or relating to, sarcoidosis. In certain embodiments, the placental cells are administered in combination with a matrix, e.g., an injectable matrix. In certain other embodiments, placental cells are administered to an individual having a sarcoidosis-related disease or disorder in combination with a solid matrix, e.g., a bone substitute, a matrix or bone substitute described in Section 4.7.4, below.
[0039] In another specific embodiment of the methods described above, said isolated placental cells are administered by bolus injection. In another specific embodiment, said isolated placental cells are administered by intravenous infusion. In a specific embodiment, said intravenous infusion is intravenous infusion over about 1 to about 8 hours. In another specific embodiment, said isolated placental cells are administered intracranially. In another specific embodiment, said isolated placental cells are administered intraperitoneally. In another specific embodiment, said isolated placental cells are administered intra-arterially. In another specific embodiment of the method of treatment, said isolated placental cells are administered intramuscularly, intradermally, subcutaneously, or intraocularly.
[0040] In another embodiment of the methods described above, said isolated placental cells are administered by surgical implantation into said individual of a composition of matter comprising said isolated placental cells. In a specific embodiment, said composition of matter is a matrix or scaffold. In another specific embodiment, said matrix or scaffold is a hydrogel. In another specific embodiment, said matrix or scaffold is a decellularized tissue.
In another specific embodiment, said matrix or scaffold is a synthetic biodegradable composition. In another specific embodiment, said matrix or scaffold is a foam.
[0041] In another specific embodiment of the methods described above, said isolated placental cells are administered once to said individual. In another specific embodiment, said isolated placental cells are administered to said individual in two or more separate administrations. In another specific embodiment, said administering comprises administering between about 1 x 104 and 1 x 105 isolated placental cells, e.g., placental cells per kilogram of said individual. In another specific embodiment, said administering comprises administering between about 1 x 105 and 1 x 106 isolated placental cells per kilogram of said individual. In another specific embodiment, said administering comprises administering between about 1 x 106 and 1 x 107 isolated placental cells per kilogram of said individual. In another specific embodiment, said administering comprises administering between about 1 x 107 and 1 x 108 isolated placental cells per kilogram of said individual. In other specific embodiments, said administering comprises administering between about 1 x 106 and about 2 x 106 isolated placental cells per kilogram of said individual; between about 2 x 106 and about 3 x 106 isolated placental cells per kilogram of said individual; between about 3 x 106 and about 4 x 106 isolated placental cells per kilogram of said individual; between about 4 x 106 and about 5 x 106 isolated placental cells per kilogram of said individual; between about 5 x 106 and about 6 x 106 isolated placental cells per kilogram of said individual; between about 6 x 106 and about 7 x 106 isolated placental cells per kilogram of said individual;
between about 7 x 106 and about 8 x 106 isolated placental cells per kilogram of said individual;
between about 8 x 106 and about 9 x 106 isolated placental cells per kilogram of said individual; or between about 9 x 106 and about 1 x 107 isolated placental cells per kilogram of said individual. In another specific embodiment, said administering comprises administering between about 1 x 107 and about 2 x 107 isolated placental cells per kilogram of said individual to said individual. In another specific embodiment, said administering comprises administering between about 1.3 x 107 and about 1.5 x 107 isolated placental cells per kilogram of said individual to said individual. In another specific embodiment, said administering comprises administering up to about 3 x 107 isolated placental cells per kilogram of said individual to said individual. In a specific embodiment, said administering comprises administering between about 5 x 106 and about 2 x 107 isolated placental cells to said individual. In another specific embodiment, said administering comprises administering about 150 x 106 isolated placental cells in about 20 milliliters of solution to said individual.
[0042] In a specific embodiment, said administering comprises administering between about x 106 and about 2 x 107 isolated placental cells to said individual, wherein said cells are contained in a solution comprising 10% dextran, e.g., dextran-40, 5% human serum albumin, and optionally an immunosuppressant.
[0043] In another specific embodiment, said administering comprises administering between about 5 x 107 and 3 x 109 isolated placental cells intravenously. In specific embodiments, said administering comprises administering about 9 x 108 isolated placental cells or about 1.8 x 109 isolated placental cells intravenously. In another specific embodiment, said administering comprises administering between about 5 x 107 and 1 x 108 isolated placental cells intralesionally. In another specific embodiment, said administering comprises administering about 9 x 107 isolated placental cells intralesionally.

3.1 Definitions [0044] As used herein, the term "about," when referring to a stated numeric value, indicates a value within plus or minus 10% of the stated numeric value.
[0045] As used herein, the term "SH2" refers to an antibody that binds an epitope on the cellular marker CD105. Thus, cells that are referred to as SH2+ are CD105+.
[0046] As used herein, the terms "SH3" and SH4" refer to antibodies that bind epitopes present on the cellular marker CD73. Thus, cells that are referred to as SH3+
and/or SH4+ are CD73+.
[0047] A placenta has the genotype of the fetus that develops within it, but is also in close physical contact with maternal tissues during gestation. As such, as used herein, the term "fetal genotype" means the genotype of the fetus, e.g., the genotype of the fetus associated with the placenta from which particular isolated placental cells, as described herein, are obtained, as opposed to the genotype of the mother that carried the fetus. As used herein, the term "maternal genotype" means the genotype of the mother that carried the fetus, e.g., the fetus associated with the placenta from which particular isolated placental cells, as described herein, are obtained.
[0048] As used herein, the term "granuloma" refers to a ball-like collection of immune cells formed as a result of inflammatory reaction, for instance, when the immune system attempts to eliminate substances (e.g., infectious organisms such as bacteria and fungi as well as other materials such as keratin, suture fragments and vegetable particles) that it perceives as foreign but is unable to eliminate.
[0049] As used herein, the term "isolated cell," e.g., "isolated placental cell," "isolated placental stem cell," and the like, means a cell that is substantially separated from other, different cells of the tissue, e.g., placenta, from which the stem cell is derived. A cell is "isolated" if at least 50%, 60%, 70%, 80%, 90%, 95%, or at least 99% of the cells, e.g., non-stem cells, with which the stem cell is naturally associated, or stem cells displaying a different marker profile, are removed from the stem cell, e.g., during collection and/or culture of the stem cell.
[0050] As used herein, "multipotent," when referring to a cell, means that the cell has the ability to differentiate into some, but not necessarily all, types of cells of the body, or into cells having characteristics of some, but not all, types of cells of the body, or into cells of one or more of the three germ layers. In certain embodiments, for example, an isolated placental cell (PDAC), as described in Section 4.2, below, that has the capacity to differentiate into a cell having characteristics of neurogenic, chondrogenic and/or osteogenic cells is a multipotent cell.
[0051] As used herein, the term "population of isolated cells" means a population of cells that is substantially separated from other cells of a tissue, e.g., placenta, from which the population of cells is derived.
[0052] As used herein, the term "placental cell" refers to a stem cell or progenitor cell that is isolated from a mammalian placenta, e.g., as described in Section 4.2, below, or cultured from cells isolated from a mammalian placenta, also referred to herein as "PDACs," either as a primary isolated or a cultured cell, regardless of the number of passages after a primary culture. In certain embodiments, the term "placental cells" as used herein does not, however, refer to, and the placental cells used in the methods provided herein are not, however, trophoblasts, cytotrophoblasts, syncitiotrophoblasts, angioblasts, hemangioblasts, embryonic germ cells, embryonic stem cells, cells obtained from an inner cell mass of a blastocyst, or cells obtained from a gonadal ridge of a late embryo, e.g., an embryonic germ cell. A cell is considered a "stem cell" if the cell displays attributes of a stem cell, e.g., a marker or gene expression profile associated with one or more types of stem cells; the ability to replicate at least 10-40 times in culture, and the ability to differentiate into cells displaying characteristics of differentiated cells of one or more of the three germ layers. Unless otherwise noted herein, the term "placental" includes the umbilical cord. The isolated placental cells disclosed herein, in certain embodiments, differentiate in vitro under differentiating conditions, differentiate in vivo, or both.
[0053] As used herein, a placental cell is "positive" for a particular marker when that marker is detectable above background. Detection of a particular marker can, for example, be accomplished either by use of antibodies, or by oligonucleotide probes or primers based on the sequence of the gene or mRNA encoding the marker. For example, a placental cell is positive for, e.g., CD73 because CD73 is detectable on placental cells in an amount detestably greater than background (in comparison to, e.g., an isotype control). A cell is also positive for a marker when that marker can be used to distinguish the cell from at least one other cell type, or can be used to select or isolate the cell when present or expressed by the cell. In the context of, e.g., antibody-mediated detection, "positive," as an indication a particular cell surface marker is present, means that the marker is detectable using an antibody, e.g., a fluorescently-labeled antibody, specific for that marker;
"positive" also refers to a cell exhibiting the marker in an amount that produces a signal, e.g., in a cytometer, that is detestably above background. For example, a cell is "CD200+" where the cell is detestably labeled with an antibody specific to CD200, and the signal from the antibody is detestably higher than that of a control (e.g., background or an isotype control). Conversely, "negative" in the same context means that the cell surface marker is not detectable using an antibody specific for that marker compared a control (e.g., background or an isotype control). For example, a cell is "CD34-" where the cell is not reproducibly detectably labeled with an antibody specific to CD34 to a greater degree than a control (e.g., background or an isotype control). Markers not detected, or not detectable, using antibodies are determined to be positive or negative in a similar manner, using an appropriate control. For example, a cell or population of cells can be determined to be OCT-4+ if the amount of OCT-4 RNA detected in RNA from the cell or population of cells is detestably greater than background as determined, e.g., by a method of detecting RNA such as RT-PCR, slot blots, etc. Unless otherwise noted herein, cluster of differentiation ("CD") markers are detected using antibodies. In certain embodiments, OCT-4 is determined to be present, and a cell is "OCT-4+" if OCT-4 is detectable using RT-PCR.
[0054] As used herein, the designation "low," when referring to the expression of a marker detectable in flow cytometry, means that the marker is expressed by fewer than 10% of cells tested, or that fluorescence attributable to the marker in, e.g., flow cytometry, is less than 1 log above background.
[0055] As used herein, "treat" encompasses the remediation of, improvement of, lessening of the severity of, or reduction in the time course of, a disease, disorder or condition (e.g,.
sarcpodpsos), or any parameter or symptom thereof.

4. DETAILED DESCRIPTION

Claims (25)

1. A method of treating an individual having sarcoidosis or a sarcoidosis-related disease or disorder, comprising administering to the individual a therapeutically effective amount of placental stem cells, or culture medium conditioned by placental stem cells, wherein the therapeutically effective amount is an amount sufficient to cause a detectable improvement in one or more symptoms of said sarcoidosis or said disease or disorder.
2. The method of claim 1, wherein said placental stem cells are CD10+, CD34-, CD105+, CD200+ placental stem cells.
3. The method of claim 2, wherein said placental stem cells are additionally CD45- or CD90+.
4. The method of claim 2, wherein said placental stem cells are additionally one or more of CD38-, CD45-, CD80-, CD86-, CD133-, HLA-DR,DP,DQ-, SSEA3-, SSEA4-, CD29+, CD44+, CD73+, CD90+, CD105+, HLA-A,B,C+, PDL1+, ABC-p+, or OCT-4+.
5. The method of claim 2, wherein said placental stem cells are additionally one or more of CD29+, CD38-, CD44+, CD54+, SH3+ or SH4+.
6. The method of claim 2, wherein said placental stem cells are additionally CD44+.
7. The method of claim 1, wherein said placental stem cells express CD200 and do not express HLA-G, or express CD73, CD105, and CD200, or express CD200 and OCT-4, or express CD73 and CD105 and do not express HLA-G, or express CD73 and CD105 and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising said stem cell when said population is cultured under conditions that allow for the formation of an embryoid-like body, or express OCT-4 and facilitate the formation of one or more embryoid-like bodies in a population of placental cells comprising said stem cell when said population is cultured under conditions that allow for the formation of an embryoid-like body.
8. The method of claim 1, wherein said sarcoidosis is systemic sarcoidosis.
9. The method of claim 8, wherein said symptom is one or more of formation of granulomas, fatigue, weight loss, fever, aches, pains, arthritis, dry eyes, swelling of the knees, blurry vision, shortness of breath, cough and skin lesions.
10. The method of claim 1, wherein said sarcoidosis is cutaneous sarcoidosis.
11. The method of claim 10, wherein the cutaneous sarcoidosis comprises annular sarcoidosis, erythrodermic sarcoidosis, ichthyosiform sarcoidosis, hypopigmented sarcoidosis, morpheaform sarcoidosis, mucosal sarcoidosis, papular sarcoid, scar sarcoid, subcutaneous sarcoidosis or ulcerative sarcoidosis.
12. The method of claim 11, wherein said symptom is a skin lesion.
13. The method of claim 12, wherein the skin lesion is one or more of erythema nodosum, nummular eczema, erythema multiforme, calcinosis cutis and pruritus, lupus pernio, skin plaques, maculopapular eruptions, nodular lesions deeper in the skin or infiltration of old scars.
14. The method of claim 1, wherein said sarcoidosis is Löfgren's syndrome.
15. The method of claim 14, wherein said symptom is one or more of erythema nodosum, bilateral hilar denopathy, arthritis, arthralgias or fever.
16. The method of claim 1, wherein said sarcoidosis is neurosarcoidosis.
17. The method of claim 16, wherein said symptom is one or more of formation of granulomas in the nervous system, headache, confusion, malaise or facial paralysis.
18. The method of claim 1, wherein said sarcoidosis is pulmonary sarcoidosis.
19. The method of claim 18, wherein said symptom is one or more of cough, shortness of breath, chest pain, a feeling of tightness in the chest, hoarseness, nasal obstruction and recurrent or persistent sinusitis.
20. The method of claim 1, wherein said sarcoidosis is cardiac sarcoidosis.
21. The method of claim 20, wherein said symptom is one or more of chest pain, palpitations, congestive heart failure, pericarditis or papillary muscle dysfunction, shortness of breath, ankle swelling, irregular heart beat or sudden death.
22. The method of claim 1, wherein said sarcoidosis is ocular sarcoidosis.
23. The method of claim 22, wherein said symptom is one or more of red or watery eyes, granulomatous uvetis, iris nodules, retinochoroiditis, conjunctivitis, lacrimal gland involvement or proptosis.
24. The method of any of claims 1-23, comprising administering a second therapeutic agent to said individual.
25. The method of claim 24, wherein said second therapeutic agent is an anti-inflammatory agent, a steroid, an immunosuppressant compound, or an antibiotic.
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