Abstract

This proposal will explore the possibility that patients with osteogenesis imperfecta (OI) can potentially be treated with their own osteoprogenitors by gene-engineening the cells from their bone marrow that are referred to as mesenchymal stem cells or marrow stromal cells (MSCs). We will use antibodies to surface epitopes we have recently identified to prepare clonal and homogeneous preparations of the earliest progenitors in cultures of MSCs that have an enhanced ability to undergo multilineage differentiation and that are rapidly self-renewing cells (RS cells). The RS cells will be used to test the hypothesis that they are similar to CD34 positive stem cells used for bone marrow transplants and that therefore they will be the most effective cells to provide long-term engraftment of osteoprogenitors into bone. We will also test the hypothesis that RS cells from patients with OI can be gene-engineered to correct the deleterious effect, of mutations in type I Collagen that produce the disease.
The Specific Aims are (1) Use a series of antibodies to surface epitopes we have recently identified to prepare clonal and homogeneous preparation, of RS cells from cultures of human MSCs. In the process, test the hypothesis that RS cells can be further fractionated to obtain homogeneous preparations of stem cells that are even more effective as osteoprogenitors for engrafment into bone. (2) Define the osteogenic potential in vitro of the RS cell preparations by assays of the rates of mineralization and assays of expressed genes by mRNA microarrays and proteomics. (3) Determine the osteogenic potential in vivo of the RS cell preparations by assays of differentiation into bone after subcutaneous implantation in vehicles or after systemic infusion into immunodeficient mice. (4) Determine the feasibility of correcting the gene defect in a patients own RS cells, (a) by overexpression of a cDNA for the wildtype COL I A I gene, or (b) by replacing a mutated COLlAl gene by homologous recombination.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR048323-04
Application #
6796230
Study Section
Special Emphasis Panel (ZAR1-TAS-C (O2))
Program Officer
Sharrock, William J
Project Start
2001-09-21
Project End
2006-07-31
Budget Start
2004-08-01
Budget End
2005-07-31
Support Year
4
Fiscal Year
2004
Total Cost
$371,250
Indirect Cost

  Institution

Name
Tulane University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
053785812
City
New Orleans
State
LA
Country
United States
Zip Code
70118

  Publications

Oskowitz, Adam Z; Penfornis, Patrice; Tucker, Alan et al. (2011) Drosha regulates hMSCs cell cycle progression through a miRNA independent mechanism. Int J Biochem Cell Biol 43:1563-72
Oskowitz, Adam; McFerrin, Harris; Gutschow, Miriam et al. (2011) Serum-deprived human multipotent mesenchymal stromal cells (MSCs) are highly angiogenic. Stem Cell Res 6:215-25
Sanchez, Cecilia G; Penfornis, Patrice; Oskowitz, Adam Z et al. (2011) Activation of autophagy in mesenchymal stem cells provides tumor stromal support. Carcinogenesis 32:964-72
Sanchez, Cecilia; Oskowitz, Adam; Pochampally, Radhika R (2009) Epigenetic reprogramming of IGF1 and leptin genes by serum deprivation in multipotential mesenchymal stromal cells. Stem Cells 27:375-82
Oskowitz, Adam Z; Lu, Jun; Penfornis, Patrice et al. (2008) Human multipotent stromal cells from bone marrow and microRNA: regulation of differentiation and leukemia inhibitory factor expression. Proc Natl Acad Sci U S A 105:18372-7
Spees, Jeffrey L; Whitney, Mandolin J; Sullivan, Deborah E et al. (2008) Bone marrow progenitor cells contribute to repair and remodeling of the lung and heart in a rat model of progressive pulmonary hypertension. FASEB J 22:1226-36
Hung, Shih-Chieh; Pochampally, Radhika R; Hsu, Shu-Ching et al. (2007) Short-term exposure of multipotent stromal cells to low oxygen increases their expression of CX3CR1 and CXCR4 and their engraftment in vivo. PLoS One 2:e416
Pochampally, Radhika R; Ylostalo, Joni; Penfornis, Patrice et al. (2007) Histamine receptor H1 and dermatopontin: new downstream targets of the vitamin D receptor. J Bone Miner Res 22:1338-49
Tolar, Jakub; Nauta, Alma J; Osborn, Mark J et al. (2007) Sarcoma derived from cultured mesenchymal stem cells. Stem Cells 25:371-9
Spees, Jeffrey L; Olson, Scott D; Whitney, Mandolin J et al. (2006) Mitochondrial transfer between cells can rescue aerobic respiration. Proc Natl Acad Sci U S A 103:1283-8

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