Compositions of placentally-derived stem cells for the treatment of cancer

Abstract

Disclosed are preparations of placentally-derived stem cells and compositions useful for the treatment of cancer. Said stem cells and compositions function through inducing a “guided differentiation” program in cancer cells, thereby reducing malignancy. Further extension of the invention pertains to augmenting ability of administered cells to induce differentiation through the co-administration of known differentiation inducing agents. Within the context of this disclosure, methods for inducing host responses to cancer are also described.

1 . A composition comprising placentally-derived tissue extract and cells, capable of inducing the differentiation of a substantially neoplastic cell into a cell with benign characteristics.  
   
   
       2 . The composition of  claim 1 , wherein the placental tissue is human.  
   
   
       3 . The composition of  claim 1 , wherein said neoplastic cell possesses one or more of the characteristics selected from the following: invasiveness, ability to metastasize, ability to suppress immune response, ability to proliferate past the Hayflick limit.  
   
   
       4 . The composition of  claim 3 , wherein said neoplastic cell is selected from the group of a cancer consisting of: a soft tissue sarcoma, a lymphoma, a cancer of the brain, an esophageal cancer, a uterine cancer, a cancer of the cervix, a bone cancer, a lung cancer, a cancer of the endometrium, a bladder cancer, a breast cancer, a cancer of the larynx, a cancer of the colon/rectum, a stomach cancer, a cancer of the ovary, a pancreatic cancers, an adrenal gland cancer and a prostate cancer.  
   
   
       5 . The composition of  claim 1 , wherein said cell with benign characteristics lacks one or more of the following: invasiveness, ability to metastasize, ability to suppress immune response, and ability to proliferate past the Hayflick limit.  
   
   
       6 . The composition of  claim 1 , wherein the placental tissue is processed through collagenase or other cell dissociating methods, followed by purification of single cells.  
   
   
       7 . The composition of  claim 6 , wherein said purification of single cells is performed using Percoll gradient of 0.3 mg/ml or other density gradients.  
   
   
       8 . The composition of  claim 1 , wherein said placentally-derived cells are cord blood stem cells.  
   
   
       9 . The composition of  claim 1 , further comprising a compound capable of inducing cellular differentiation.  
   
   
       10 . The composition of  claim 9 , wherein said compound capable of inducing cellular differentiation is selected from the group consisting of: dihydroxyvitamin D3, retinoic acid, or valproic acid.  
   
   
       11 . The composition of  claim 9 , wherein said compound capable of inducing cellular differentiation is a histone deacetylase inhibitor.  
   
   
       12 . The composition of  claim 11 , wherein said histone deacetylase inhibitor is selected from the group consisting of: trichostatin A, a short-chain fatty acid, a hydroxamate, a cyclic tetrapeptide, or a benzamide.  
   
   
       13 . The composition of  claim 1 , wherein said composition is an adjuvant to a chemotherapeutic drug.  
   
   
       14 . A method of reducing malignancy of a tumor in a patient comprising: 
 a. extracting placental tissue obtained from a healthy mother;    b. homogenizing said tissue and obtaining a single cell suspension;    c. purifying CD34+ cells; and    d. administering the CD34+ cells to a patient in need thereof.    
   
   
       15 . The method of  claim 14 , wherein a differentiation inducing agent is co-administered with the CD34+ cells.  
   
   
       16 . The method of  claim 14 , wherein said CD34+ cells are expanded in vitro prior to administration.  
   
   
       17 . The method of  claim 16 , wherein expansion is performed by culture in media conditioned with cytokines and/or growth factors selected from the group consisting of: leukemia inhibitory factor, IL-1 through IL-13, IL-15 through IL-17, IL-19 through IL-22, granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), erythropoietin (Epo), thrombopoietin (Tpo), Flt3-ligand, B cell activating factor, artemin, bone morphogenic protein factors, epidermal growth factor (EGF), glial derived neurotrophic factor, lymphotactin, macrophage inflammatory proteins, myostatin, neurturin, nerve growth factors, platelet derived growth factors, placental growth factor, pleiotrophin, stern cell factor, stem cell growth factors, transforming growth factors, tumor necrosis factors, Vascular Endothelial Cell Growth Factors, and fibroblast growth factors, FGF-acidic and basic fibroblast growth factor.  
   
   
       18 . The method of  claim 14 , wherein one or more genes is transfected into the CD34+ cells prior to administration.  
   
   
       19 . The method of  claim 18 , wherein said one or more transfected genes are associated with inhibition of tumor growth.  
   
   
       20 . The method of  claim 19 , wherein said one or more transfected genes are selected from the group consisting of: IFN-g, IL-2, IL12, IL-15, IL-18, IL-23.  
   
   
       21 . The method of  claim 14 , wherein a chemotherapeutic agent is co-administered with the CD34+ cells.  
   
   
       22 . The method of  claim 21 , wherein said chemotherapeutic agent is selected from the group consisting of: methotrexate, vincristine, adriamycin, cisplatin, non-sugar containing chloroethylnitrosoureas, 5-fluorouracil, mitomycin C, bleomycin, doxorubicin, dacarbazine, taxol, fragyline, Meglamine GLA, valrubicin, carmustaine and poliferposan, MM1270, BAY 12-9566, RAS famesyl transferase inhibitor, famesyl transferase inhibitor, MMP, MTA/LY231514, LY264618/Lometexol, Glamolec, CI-994, TNP-470, Hycamtin/Topotecan, PKC412, Valspodar/PSC833, Novantrone/Mitroxantrone, Metaret/Suramin, Batimastat, E7070, BCH-4556, CS-682, 9-AC, AG3340, AG3433, Incel/VX-710, VX-853, ZD0101, ISI641, ODN 698, TA 2516/Marmistat, BB2516/Marmistat, CDP 845, D2163, PD183805, DX8951f, Lemonal DP 2202, FK 317, Picibanil/OK-432, AD 32/Valrubicin, Metastron/strontium derivative, Temodal/Temozolomide, Evacet/liposomal doxorubicin, Yewtaxan/Placlitaxel, Taxol/Paclitaxel, Xeload/Capecitabine, Furtulon/Doxifluridine, Cyclopax/oral paclitaxel, Oral Taxoid, SPU-077/Cisplatin, HMR 1275/Flavopiridol, CP-358 (774)/EGFR, CP-609 (754)/RAS oncogene inhibitor, BMS-182751/oral platinum, UFT (Tegafur/Uracil), Ergamisol/Levamisole, Eniluracil/776C85/5FU enhancer, Campto/Levamisole, Camptosar/Irinotecan, Tumodex/Ralitrexed, Leustatin/Cladribine, Paxex/Paclitaxel, Doxil/liposomal doxorubicin, Caelyx/liposomal doxorubicin, Fludara/Fludarabine, Pharmarubicin/Epirubicin, DepoCyt, ZD1839, LU 79553/Bis-Naphtalimide, LU 103793/Dolastain, Caetyx/liposomal doxorubicin, Gemzar/Gemcitabine, ZD 0473/Anormed, YM 116, Iodine seeds, CDK4 and CDK2 inhibitors, PARP inhibitors, D4809/Dexifosamide, Ifes/Mesnex/Ifosamide, Vumon/Teniposide, Paraplatin/Carboplatin, Plantinol/cisplatin, Vepeside/Etoposide, ZD 9331, Taxotere/Docetaxel, prodrug of guanine arabinoside, Taxane Analog, nitrosoureas, alkylating agents such as melphelan and cyclophosphamide, Aminoglutethimide, Asparaginase, Busulfan, Carboplatin, Chlorombucil, Cytarabine HCl, Dactinomycin, Daunorubicin HCl, Estramustine phosphate sodium, Etoposide (VP16-213), Floxuridine, Fluorouracil (5-FU), Flutamide, Hydroxyurea (hydroxycarbamide), Ifosfamide, Interferon Alfa-2a, Alfa-2b, Leuprolide acetate (LHRH-releasing factor analogue), Lomustine (CCNU), Mechlorethamine HCl (nitrogen mustard), Mercaptopurine, Mesna, Mitotane (o.p′-DDD), Mitoxantrone HCl, Octreotide, Plicamycin, Procarbazine HCl, Streptozocin, Tamoxifen citrate, Thioguanine, Thiotepa, Vinblastine sulfate, Amsacrine (m-AMSA), Azacitidine, Erthropoietin, Hexamethylmelamine (HMM), Interleukin 2, Mitoguazone (methyl-GAG; methyl glyoxal bis-guanylhydrazone; MGBG), Pentostatin (2′deoxycoformycin), Semustine (methyl-CCNU), Teniposide (VM-26) and Vindesine sulfate.  
   
   
       23 . A cell free extract capable of inducing differentation in cancer cells, the cell free extract produced by the process comprising: 
 a. extracting human placenta;    b. homogenizing said placenta, resulting in a homogenate;    C. centrifuging said homogenate in order to free the supernatant of cellular debris;    d. purifying differentiation associated molecular weight fractions; and    e. administering said fractions to a patient in need thereof.    
   
   
       24 . The cell free extract of  claim 23  wherein the differentiation-inducing fraction purified is of molecular weight of 10-10,000 Daltons.  
   
   
       25 . The cell free extract of  claim 23  wherein the differentiation-inducing fraction purified is of molecular weight of 300-3,000 Daltons.  
   
   
       26 . A method of stimulating an immune response to cancer cells by administration of the cell free extract of  claim 23 , together with an appropriate immunological adjuvant.  
   
   
       27 . A method of prophylactically treating a cancer patient by administration of the cell free extract of  claim 23  to a patient in need thereof.  
   
   
       28 . A method of inhibiting progression of a preneoplastic lesion to neoplasia by administration of the cell free extract of  claim 23 .  
   
   
       29 . A cell free extract capable of inducing differentiation in cancer cells, the cell free extract produced by the process comprising: 
 a. extracting bone marrow stem cells or unpurified bone marrow nucleated cells;    b. homogenizing said cells, resulting in a homogenate;    c. centrifuging said homogenate in order to free the supernatant of cellular debris;    d. purifying differentiation associated molecular weight fractions; and    e. administering said fractions to a patient in need thereof.    
   
   
       30 . A differentiation inducing composition, suitable for administration to a patient in need thereof, derived from placental tissue, the composition produced by the process comprising: 
 a. extracting human placenta;    b. homogenizing said placenta, resulting in a homogenate;    c. centrifuging said homogenate in order to free the supernatant of cellular debris;    d. purifying the cell free-debris through adsorption on a C-18 column; and    e. filter sterilizing said composition.    
   
   
       31 . The composition of  claim 30 , wherein the homogenized portion of the placenta consists substantially of chorionic tissue.  
   
   
       32 . The composition of  claim 30 , wherein a histone deacetylase inhibitor is added directly to the composition.  
   
   
       33 . A method of inducing differentiation and reprogramming of distal cancer lesions through an intratumoral administration of the composition described in  claim 30 .  
   
   
       34 . A method of substantially increasing efficacy of the composition of  claim 30  by administration in a liposomal complex.  
   
   
       35 . A differentiation inducing composition, comprising a substantially purified therapeutic composition prepared through the following steps: 
 (a) preparing a supernatant;    (b) subjecting the supernatant to solid phase extraction to produce a product;    (c) subjecting the product from step (b) to gel filtration to produce a product;    (d) subjecting the product from step (c) to anion exchange fast phase liquid chromatography to produce a product;    (e) subjecting the product from step (d) to amino-high performance liquid chromatography to produce a product; and    (f) subjecting the product from step (e) to reverse phase high performance liquid chromatography to produce purified differentiation inducing factor.    
   
   
       36 . The composition of  claim 35 , wherein said supernatant is prepared by culturing cord-blood derived stem cells to produce a factor-containing supernatant.  
   
   
       37 . The composition of  claim 35 , wherein said supernatant is prepared through centrifugation of lysed placental extract.