Methods and compositions for treatment of bone defects with placental cell populations

• US 8,728,805 B2
• Filed: 08/24/2009
• Issued: 05/20/2014
• 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, Heidaran, Mohammad A., Labazzo, Kristen, Yaccoby, Shmuel
• Primary Examiner(s): Weber, Jon P
• Assistant Examiner(s): Knight, Teresa E
• Application Number:
  US12/546,556
• Publication Number:
  US 20100047214A1
• Time in Patent Office: 1,730 Days
• Field of Search: None
• US Class Current: 435/325
• 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 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 an isolated population of cells comprising osteogenic placental adherent cells (OPACs),

wherein said OPACs are obtained from chorion, are adherent to tissue culture plastic, are negative for CD200 or are CD200^dim, and positive for CD 105,

wherein at least 80% of the cells of said population are OPACs, 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⁺ adherent 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 CD200⁺ adherent 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 CD200⁺ adherent 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 CD200⁺ adherent 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 CD200⁺ adherent 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 CD200⁺ adherent 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 said bone marrow-derived 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 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 said bone marrow-derived mesenchymal 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 bone marrow-derived mesenchymal 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 said bone marrow-derived 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 said bone marrow-derived mesenchymal 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 bone marrow-derived mesenchymal 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 bone marrow-derived 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 OPACs and 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.

10. A pharmaceutical composition comprising an isolated population of cells comprising osteogenic placental adherent cells (OPACs), wherein said OPACs are obtained from chorion, adherent to tissue culture plastic, negative for CD200 or are CD200^dim, and positive for CD105, and wherein at least 80% of the cells of said population are OPACs.

11. The pharmaceutical composition of claim 10, 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⁺ adherent 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 CD200⁺ adherent 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 CD200⁺ adherent 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 CD200⁺ adherent 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 CD200⁺ adherent 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 CD200⁺ adherent 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 said bone marrow-derived 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 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 said bone marrow-derived mesenchymal 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 bone marrow-derived mesenchymal 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 said bone marrow-derived 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 said bone marrow-derived mesenchymal 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 bone marrow-derived mesenchymal stem cells are cultured in growth medium or osteogenic medium, as assessed by Ct values from quantitative real-time PCR.

12. The pharmaceutical composition of claim 10, wherein said OPACs 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.

13. The pharmaceutical composition of claim 10, wherein said OPACs 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.

14. The pharmaceutical composition of claim 10, wherein at least 95% of the cells of said population are OPACs.

15. The pharmaceutical composition of claim 10, wherein at least 99% of the cells of said population are OPACs.

16. The pharmaceutical composition of claim 10, wherein said OPACs are SSEA3⁺ or SSEA4⁺.

17. The pharmaceutical composition of claim 10, wherein said OPACs are SSEA3⁺ and SSEA4⁺.

18. The pharmaceutical composition of claim 10, wherein said OPACs produce osteoprotegerin.

19. The pharmaceutical composition of claim 10, wherein said OPACs are 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 exhibit inducible alkaline phosphatase activity.

20. The pharmaceutical composition of claim 10, wherein said OPACs are 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 exhibit inducible alkaline phosphatase activity.