An attractive modality for bone regeneration involves the use of pluripotent mesenchymal stem cells that are induced by osteogenic factors. Our overall goal is to investigate differential effects of bone morphogenetic proteins on bone healing in murine craniofacial defect models. This project is based on the hypothesis that there is a definable, differential profile of the osteogenic BMPs (BMP2, 4, 6, 7, 9) in regards to their capacity to induce healing of critical-sized craniofacial defects. To test this hypothesis, the following specific aims are proposed: 1) i. To construct recombinant adenoviruses expressing individual BMPs (BMP-2, -4, -6, -7, -9), ii. To determine the effect of stem cell transfection with the recombinant adenoviruses above on the expression levels of various proliferation and/or osteogenic differentiation markers in the stem cells, iii. To determine the differential effects of the various BMPs (2, 4, 6, 7, 9) on bone healing of critical sized defects in murine models. iv. To characterize this differential effect in areas of non-load bearing (calvarial) and areas of load-bearing (mandibular) craniofacial repair;2) v. To generate primary cultures of calvarial/mandibular osteoblasts for in vitro stimulation with BMP, vi.To stimulate primary cultures of isolated osteoblasts with the various BMPs and measure the capacity of each BMP to induce osteogenesis in early and late stages of differentiation;3) vii. To target canonical Wnt/beta catenin signaling via si-RNA technology and examine the effects of pathway inhibition both in vitro and in vivo. viii. To target SMAD4 signaling via si-RNA technology and examine the effects of SMAD4 inhibition both in vitro and in vivo.
These specific aims will be addressed by the following experimental design: 1) Transfection of mesenchymal stem cells with adenovirus encoded with BMPs (Ad-BMPs);2) Infection of mesenchymal stem cells with Ad-BMPs and testing of these stem cells in vitro for osteogenic differentiation;3) Transfer of Ad-BMP induced mesenchymal stem cells into critical-sized murine craniofacial defects and evaluation of defect closure via micro-CT;4) RT-PCR of downstream signalling elements important in BMP-induced stem cell osteogenesis;5) siRNA knockdown of these critical signalling elements both in vitro and in vivo. Public Health Relevance: Large defects from birth, trauma or cancer resection require almost a limitless source of bone, which cannot be supplied by autogenous donor sites without serious morbidity. A means of providing large quantities of bone would be to transfer engineered, pluripotent stem cells that could induce bony regeneration. The project characterized herein has relevance to patients with skeletal deficiencies of the craniofacial skeleton.

Public Health Relevance

Large defects from birth, trauma or cancer resection require almost a limitless source of bone, which cannot be supplied by autogenous donor sites without serious morbidity. A means of providing large quantities of bone would be to transfer engineered, pluripotent stem cells that could induce bony regeneration. The project characterized herein has relevance to patients with skeletal deficiencies of the craniofacial skeleton.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Clinical Investigator Award (CIA) (K08)
Project #
1K08DE020140-01
Application #
7774491
Study Section
NIDCR Special Grants Review Committee (DSR)
Program Officer
Hardwick, Kevin S
Project Start
2009-09-25
Project End
2014-08-31
Budget Start
2009-09-25
Budget End
2010-08-31
Support Year
1
Fiscal Year
2009
Total Cost
$123,550
Indirect Cost
Name
University of Chicago
Department
Surgery
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
Shu, Yi; Yang, Chao; Ji, Xiaojuan et al. (2018) Reversibly immortalized human umbilical cord-derived mesenchymal stem cells (UC-MSCs) are responsive to BMP9-induced osteogenic and adipogenic differentiation. J Cell Biochem 119:8872-8886
Zeng, Zongyue; Huang, Bo; Huang, Shifeng et al. (2018) The development of a sensitive fluorescent protein-based transcript reporter for high throughput screening of negative modulators of lncRNAs. Genes Dis 5:62-74
Hu, Xue; Li, Li; Yu, Xinyi et al. (2017) CRISPR/Cas9-mediated reversibly immortalized mouse bone marrow stromal stem cells (BMSCs) retain multipotent features of mesenchymal stem cells (MSCs). Oncotarget 8:111847-111865
Pakvasa, Mikhail; Alverdy, Alex; Mostafa, Sami et al. (2017) Neural EGF-like protein 1 (NELL-1): Signaling crosstalk in mesenchymal stem cells and applications in regenerative medicine. Genes Dis 4:127-137
Liao, Junyi; Yu, Xinyi; Hu, Xue et al. (2017) lncRNA H19 mediates BMP9-induced osteogenic differentiation of mesenchymal stem cells (MSCs) through Notch signaling. Oncotarget 8:53581-53601
Liao, J; Wei, Q; Fan, J et al. (2017) Characterization of retroviral infectivity and superinfection resistance during retrovirus-mediated transduction of mammalian cells. Gene Ther 24:333-341
Bishop, Elliot S; Mostafa, Sami; Pakvasa, Mikhail et al. (2017) 3-D bioprinting technologies in tissue engineering and regenerative medicine: Current and future trends. Genes Dis 4:185-195
Song, Dongzhe; Zhang, Fugui; Reid, Russell R et al. (2017) BMP9 induces osteogenesis and adipogenesis in the immortalized human cranial suture progenitors from the patent sutures of craniosynostosis patients. J Cell Mol Med 21:2782-2795
Lee, Cody S; Bishop, Elliot S; Zhang, Ruyi et al. (2017) Adenovirus-Mediated Gene Delivery: Potential Applications for Gene and Cell-Based Therapies in the New Era of Personalized Medicine. Genes Dis 4:43-63
Ye, Jixing; Wang, Jing; Zhu, Yunxiao et al. (2016) A thermoresponsive polydiolcitrate-gelatin scaffold and delivery system mediates effective bone formation from BMP9-transduced mesenchymal stem cells. Biomed Mater 11:025021

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