Methods and Compositions for Treatment of Bone Defects with Placental Cell Populations

• US 20100047214A1
• Filed: 08/24/2009
• Published: 02/25/2010
• Est. Priority Date: 08/22/2008
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Abramson, Sascha D., Guelakis, Marian, Yaccoby, Shmuel, Labazzo, Kristen, Heidaran, Mohammad A.
• Granted Patent: US 8,728,805 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly

Abstract

Provided herein are osteogenic placental adherent cells (OPACs), methods of using OPACs and OPAC populations, and methods of culturing, proliferating, expanding, or differentiating the OPACs. Further provided herein are methods of using the OPACs to formulate implantable or injectable compositions suitable for administration to a subject. Still further provided herein are provides methods for treating bone defects with OPACs and compositions comprising OPACs. Also provided herein are methods of using OPACs in the treatment and management of multiple myeloma, e.g., reducing the progression of, halting the progression of, or improving, one or more symptoms of multiple myeloma in an individual having multiple myeloma, comprising administering a plurality of OPACs to the individual.

Claims

1. A method of treating an individual having multiple myeloma, comprising administering to said individual isolated osteogenic placental adherent cells (OPACs), wherein said OPACs are obtained from chorion, and are adherent to tissue culture plastic, and wherein said OPACs are negative for CD200 or are CD200^dim, and positive for CD105, and wherein said administering detectably reduces the progression of, halts the progression of, or improves, one or more symptoms of said multiple myeloma.

2. The method of claim 1, wherein said OPACs are SSEA3⁺ or SSEA4⁺.

3. The method of claim 1, wherein said OPACs are SSEA3⁺ and SSEA4⁺.

4. The method of claim 1, wherein said OPACs:

express one or more genes at a detectably higher level than an equivalent number of CD200⁺ adherent placental stem cells, wherein said one or more genes comprise one or more of BMP3 (bone morphogenetic protein

     3), CDH11, COL10A1, COL14A1, COL15A1, DMP1 (dentin matrix acidic phosphoprotein

     1), DSPP (dentin sialophosphoprotein), ENAM (enamelin), FGFR2 (fibroblast growth factor receptor

     2), MMP10 (matrix metalloprotease 10 (stromelysin

     2)), TGFB3, and/or TGFBR1, when said OPACs and said CD200⁺ placental stem cells are cultured in growth medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of CD200⁺ adherent placental stem cells, wherein said one or more genes comprise one or more of AMBN (ameloblastin (enamel matrix protein)), BMP2 (bone morphogenetic protein

     2), CALCR (calcitonin receptor), CDH11, COL11A1, COL14A1, COL15A1, COL2A1, CSF2, CSF3, DMP1, DSPP, ENAM, FGF3, GDF10 (growth differentiation factor

     10), IGF1 (insulin-like growth factor

     1), ITGA1 (integrin, alpha 1 (CD49)), ITGA2 (integrin, alpha 2 (CD49B)), MMP10, MMP8 (matrix metalloprotease 8 (neutrophil collagenase)), MMP9, PDGFA (platelet-derived growth factor A), SMAD1, TGFB3, TGFBR1 and/or TGFBR2 (transforming growth factor beta, receptor

     2) when said OPACs and said placental stem cells are cultured in osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of adherent CD200⁺ placental stem cells that are not trophoblasts or cytotrophoblasts, wherein said one or more genes comprise one or more, or all, of CDH11, COL14A1, COL15A1, DMP1, DSPP, ENAM, MMP10, TGFB3 and/or TGFBR1 regardless of whether said OPACs and said placental stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably lower level than an equivalent number of adherent CD200⁺ placental stem cells that are not trophoblasts or cytotrophoblasts, wherein said one or more genes comprise one or more, or all, of AHSG (alpha-2-HS-glycoprotein), ALPL (alkaline phosphatase liver/bone/kidney), EGF (epidermal growth factor), FLT1 (fins-related tyrosine kinase 1 (vascular endothelial growth factor/vascular permeability factor receptor)), IGF2, ITGA2, ITGAM (integrin, alpha M (complement component 3 receptor 3 subunit)), SCARB1 (scavenger receptor class B, member

     1), SOX9 (SRY (sex determining region Y)-box

     9), TNF, TWIST1 (Twist homolog 1;

formerly blepharophimosis, epicanthus inversus and ptosis 3, acrocephalosyndactyly

     3), VCAM1 (vascular cell adhesion molecule

     1) and/or VDR (vitamin D (1,25-dihydroxyvitamin D3) receptor) when said OPACs and said placental stem cells are cultured in growth medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably lower level than an equivalent number of adherent CD200⁺ placental stem cells that are not trophoblasts or cytotrophoblasts, wherein said one or more genes comprise one or more, or all, of BGN (biglycan), COL11A1, COMP (cartilage oligomeric matrix protein), FGF1 and/or VCAM1 when said OPACs and said placental stem cells are cultured in osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express VCAM1 at a detectably lower level than an equivalent number of adherent CD200⁺ placental stem cells that are not trophoblasts or cytotrophoblasts, regardless of whether said OPACs and said placental stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of BMP4, CALCR, CD36, CDH11, COL12A1, COL14A1, COL15A1, COL3A1, COL5A1, DMP1, DSPP, FLT1, MSX1, PDGFA, TGFB3, TGFBR1 and/or TUFT1 (Tuftelin

     1), when the OPACs and stem cells are cultured in growth medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of AMBN, CALCR, COL14A1, COL15A1, CSF3, DMP1, DSPP, ITGA1, ITGA2, MMP10, MMP9, MSX1, PDGFA, TGFB1, TGFB3, TGFBR1 and/or TGFBR2, when the OPACs and stem cells are cultured in osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of CALCR, COL14A1, COL15A1, DMP1, DSPP, MSX1, PDGFA, TGFB3 and/or TGFBR1 regardless of whether said OPACs and said placental stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably lower level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of ALPL, BGLAP (bone gamma-carboxyglutamate (gla) protein), IGF2, ITGA2, ITGAM, SCARB1 and/or SOX1, when the OPACs and mesenchymal stem cells are cultured in growth medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably lower level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of AHSG, ALPL, BGLAP, BGN, BMP3, BMP5, CD36, COL10A1, COL11A1, COL12A1, COL2A1, COL4A3, COMP, EGF, FGF1, FGFR2, IGF2, MMP8, PHEX (phosphate regulating endopeptidase homolog, X-linked), RUNX2 (runt-related transcription factor

     2), SCARB1, SOX1, VCAM1 and/or VEGFB, when the OPACs and stem cells are cultured in osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

or

express one or more genes at a detectably lower level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of ALPL, BGLAP, IGF2, SCARB1 and/or SOX9, regardless of whether said OPACs and said placental stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR.

5. The method of claim 1, wherein said OPACs:

express one or more genes at a detectably higher level than an equivalent number of bone marrow derived mesenchymal stem cells, wherein said one or more genes comprise one or more of BMP4, BMP6, CD36, CDH11, COL14A1, COL15A1, COL1A1, COL3A1, COL5A1, CSF2, CTSK, FGF2, FGFR1, FLT1, ITGA1, MINPP1, MMP9, MSX1, PDGFA, SERPINH1, TGFB3 and TGFBR1, wherein said OPACs and said mesenchymal stem cells have undergone an equivalent number of passages;

or

express one or more genes at a detectably higher level than an equivalent number of fibroblast cells, wherein said one or more genes comprise one or more of BMP4, BMP6, CDH11, COL14A1, COL15A1, COL1A1, COL3A1, COL5A1, FLT1, IGF1R, ITGA1, MINPP1, PDGFA, SERPINH1, SMAD3, TGFB1, TGFB2, TGFB3, TGFBR1, TNF, TUFT1, VCAM1 and VEGFA, and wherein said fibroblast cells have undergone an equivalent number of passages.

6. The method of claim 1, wherein said OPACs secrete one or more of the proteins decorin, epiregulin, IGFBP-3, IGFBP-6, IL-2 R alpha, IL-17RC, IL-27, Latent TGF-beta binding protein 1 (LTBP), NCAM-1, Smad4, TFPI, TGF-beta R1/ALK5 or TIMP-2.

7. The method of claim 1, wherein said OPACs secrete the proteins decorin, epiregulin, IGFBP-3, IGFBP-6, IL-2 R alpha, IL-17RC, IL-27, Latent TGF-beta binding protein 1 (LTBP), NCAM-1, Smad4, TFPI, TGF-beta R1/ALK5 and TIMP-2.

8. The method of claim 1, wherein said one or more symptoms are bone pain, osteocytic lesions, anemia, or renal failure.

9. The method of claim 1, comprising administering at least 1×

10⁸ OPACs/kg to said individual.

11. The isolated cell of claim 9 that is SSEA3⁺ or SSEA4⁺.

12. The isolated cell of claim 9 that is SSEA3⁺ and SSEA4⁺.

13. The isolated cell of claim 9, wherein said cell produces osteoprotegerin.

14. The isolated cell of claim 9, wherein said cell is additionally negative for expression of α

-smooth muscle actin, negative for expression of RANKL, positive for expression of NG2, positive for expression of osteoprotegerin, or exhibits inducible alkaline phosphatase activity.

15. The isolated cell of claim 13, wherein said cell is additionally negative for expression of α

-smooth muscle actin, negative for expression of RANKL, positive for expression of NG2, positive for expression of osteoprotegerin, and exhibits inducible alkaline phosphatase activity.

16. The isolated cell of claim 9, wherein said cell:

expresses one or more genes at a detectably higher level than a CD200⁺ adherent placental stem cell, wherein said one or more genes comprise one or more of BMP6, CDH11, COL10A1, COL14A1, COL15A1, COL1A1, COL1A2, COL3A1, COL4A3, COL5A1, CSF3, CTSK, IGF1R, MINPP1, MMP2, MMP9, MSX1, SMAD1, SMAD3, TGFB3, TGFBR1 and VEGFB, wherein said OPAC and said CD200⁺ adherent placental stem cell have undergone an equivalent number of passages;

expresses one or more genes at a detectably higher level than a bone marrow derived mesenchymal stem cell, wherein said one or more genes comprise one or more of BMP4, BMP6, CD36, CDH11, COL14A1, COL15A1, COL1A1, COL3A1, COL5A1, CSF2, CTSK, FGF2, FGFR1, FLT1, ITGA1, MINPP1, MMP9, MSX1, PDGFA, SERPINH1, TGFB3 and TGFBR1, wherein said OPAC and said mesenchymal stem cell have undergone an equivalent number of passages; and

/or

expresses one or more genes at a detectably higher level than a fibroblast cell, wherein said one or more genes comprise one or more of BMP4, BMP6, CDH11, COL14A1, COL15A1, COL1A1, COL3A1, COL5A1, FLT1, IGF1R, ITGA1, MINPP1, PDGFA, SERPINH1, SMAD3, TGFB1, TGFB2, TGFB3, TGFBR1, TNF, TUFT1, VCAM1 and VEGFA, wherein said fibroblast cell has undergone a number of passages in culture that is equivalent to the number of passages said fibroblast cell has undergone.

17. The isolated cell of claim 9, wherein said cell secretes one or more of the proteins decorin, epiregulin, IGFBP-3, IGFBP-6, IL-2 R alpha, IL-17RC, IL-27, Latent TGF-beta binding protein 1 (LTBP), NCAM-1, Smad4, TFPI, TGF-beta R1/ALK5 or TIMP-2.

18. The isolated cell of claim 16, wherein said cell secretes the proteins decorin, epiregulin, IGFBP-3, IGFBP-6, IL-2 R alpha, IL-17RC, IL-27, Latent TGF-beta binding protein 1 (LTBP), NCAM-1, Smad4, TFPI, TGF-beta R1/ALK5 and TIMP-2.

19. An isolated population of cells comprising the cells of claim 9.

20. The isolated population of cells of claim 9, wherein said cells:

express one or more genes at a detectably higher level than an equivalent number of CD200⁺ adherent placental stem cells, wherein said one or more genes comprise one or more of BMP3 (bone morphogenetic protein

     3), CDH11, COL10A1, COL14A1, COL15A1, DMP1 (dentin matrix acidic phosphoprotein

     1), DSPP (dentin sialophosphoprotein), ENAM (enamelin), FGFR2 (fibroblast growth factor receptor

     2), MMP10 (matrix metalloprotease 10 (stromelysin

     2)), TGFB3, and/or TGFBR1, when said OPACs and said CD200⁺ placental stem cells are cultured in growth medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of CD200⁺ adherent placental stem cells, wherein said one or more genes comprise one or more of AMBN (ameloblastin (enamel matrix protein)), BMP2 (bone morphogenetic protein

     2), CALCR (calcitonin receptor), CDH11, COL11A1, COL14A1, COL15A1, COL2A1, CSF2, CSF3, DMP1, DSPP, ENAM, FGF3, GDF10 (growth differentiation factor

     10), IGF1 (insulin-like growth factor

     1), ITGA1 (integrin, alpha 1 (CD49)), ITGA2 (integrin, alpha 2 (CD49B)), MMP10, MMP8 (matrix metalloprotease 8 (neutrophil collagenase)), MMP9, PDGFA (platelet-derived growth factor A), SMAD1, TGFB3, TGFBR1 and/or TGFBR2 (transforming growth factor beta, receptor

     2) when said OPACs and said placental stem cells are cultured in osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of adherent CD200⁺ placental stem cells that are not trophoblasts or cytotrophoblasts, wherein said one or more genes comprise one or more, or all, of CDH11, COL14A1, COL15A1, DMP1, DSPP, ENAM, MMP10, TGFB3 and/or TGFBR1 regardless of whether said OPACs and said placental stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably lower level than an equivalent number of adherent CD200⁺ placental stem cells that are not trophoblasts or cytotrophoblasts, wherein said one or more genes comprise one or more, or all, of AHSG (alpha-2-HS-glycoprotein), ALPL (alkaline phosphatase liver/bone/kidney), EGF (epidermal growth factor), FLT1 (fms-related tyrosine kinase 1 (vascular endothelial growth factor/vascular permeability factor receptor)), IGF2, ITGA2, ITGAM (integrin, alpha M (complement component 3 receptor 3 subunit)), SCARB1 (scavenger receptor class B, member

     1), SOX9 (SRY (sex determining region Y)-box

     9), TNF, TWIST1 (Twist homolog 1;

formerly blepharophimosis, epicanthus inversus and ptosis 3, acrocephalosyndactyly

     3), VCAM1 (vascular cell adhesion molecule

     1) and/or VDR (vitamin D (1,25-dihydroxyvitamin D3) receptor) when said OPACs and said placental stem cells are cultured in growth medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably lower level than an equivalent number of adherent CD200⁺ placental stem cells that are not trophoblasts or cytotrophoblasts, wherein said one or more genes comprise one or more, or all, of BGN (biglycan), COL11A1, COMP (cartilage oligomeric matrix protein), FGF1 and/or VCAM1 when said OPACs and said placental stem cells are cultured in osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express VCAM1 at a detectably lower level than an equivalent number of adherent CD200⁺ placental stem cells that are not trophoblasts or cytotrophoblasts, regardless of whether said OPACs and said placental stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of BMP4, CALCR, CD36, CDH11, COL12A1, COL14A1, COL15A1, COL3A1, COL5A1, DMP1, DSPP, FLT1, MSX1, PDGFA, TGFB3, TGFBR1 and/or TUFT1 (Tuftelin

     1), when the OPACs and stem cells are cultured in growth medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of AMBN, CALCR, COL14A1, COL15A1, CSF3, DMP1, DSPP, ITGA1, ITGA2, MMP10, MMP9, MSX1, PDGFA, TGFB1, TGFB3, TGFBR1 and/or TGFBR2, when the OPACs and stem cells are cultured in osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably higher level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of CALCR, COL14A1, COL15A1, DMP1, DSPP, MSX1, PDGFA, TGFB3 and/or TGFBR1 regardless of whether said OPACs and said placental stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably lower level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of ALPL, BGLAP (bone gamma-carboxyglutamate (gla) protein), IGF2, ITGA2, ITGAM, SCARB1 and/or SOX1, when the OPACs and mesenchymal stem cells are cultured in growth medium, as assessed by Ct values from quantitative real-time PCR;

express one or more genes at a detectably lower level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of AHSG, ALPL, BGLAP, BGN, BMP3, BMP5, CD36, COL10A1, COL11A1, COL12A1, COL2A1, COL4A3, COMP, EGF, FGF1, FGFR2, IGF2, MMP8, PHEX (phosphate regulating endopeptidase homolog, X-linked), RUNX2 (runt-related transcription factor

     2), SCARB1, SOX1, VCAM1 and/or VEGFB, when the OPACs and stem cells are cultured in osteogenic medium, as assessed by Ct values from quantitative real-time PCR;

or

express one or more genes at a detectably lower level than an equivalent number of bone marrow-derived mesenchymal stem cells, wherein said one or more genes comprise one or more, or all, of ALPL, BGLAP, IGF2, SCARB1 and/or SOX9, regardless of whether said OPACs and said placental stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR.

21. The isolated population of cells of claim 18, wherein cells in said population of cells express one or more of the proteins decorin, epiregulin, IGFBP-3, IGFBP-6, IL-2 R alpha, IL-17RC, IL-27, Latent TGF-beta binding protein 1 (LTBP), NCAM-1, Smad4, TFPI, TGF-beta R1/ALK5 or TIMP-2.

22. The isolated population of cells of claim 18, wherein cells in said population of cells express the proteins decorin, epiregulin, IGFBP-3, IGFBP-6, IL-2 R alpha, IL-17RC, IL-27, Latent TGF-beta binding protein 1 (LTBP), NCAM-1, Smad4, TFPI, TGF-beta R1/ALK5 and TIMP-2.

23. The isolated population of cells of claim 18, wherein at least 50% of the cells of said population are the cells of claim 9.

24. The isolated population of cells of claim 18, wherein at least 80% of the cells of said population are the cells of claim 9.

25. The isolated population of cells of claim 18, wherein at least 95% of the cells of said population are the cells of claim 9.

26. The isolated population of cells of claim 18 consisting essentially of the cells of claim 9.

10. An isolated osteogenic placental adherent cell (OPAC), wherein said cell is obtained from chorion, and is adherent to tissue culture plastic, and wherein said cell is negative for CD200 or is CD200^dim, and positive for CD105.