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.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08DE020140-05
Application #
8519411
Study Section
NIDCR Special Grants Review Committee (DSR)
Program Officer
King, Lynn M
Project Start
2009-09-25
Project End
2014-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
5
Fiscal Year
2013
Total Cost
$125,166
Indirect Cost
$9,272
Name
University of Chicago
Department
Surgery
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
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Lu, Shun; Wang, Jing; Ye, Jixing et al. (2016) Bone morphogenetic protein 9 (BMP9) induces effective bone formation from reversibly immortalized multipotent adipose-derived (iMAD) mesenchymal stem cells. Am J Transl Res 8:3710-3730
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
Lyon, Sarah M; Waggoner, Darrel; Halbach, Sara et al. (2015) Syndromic craniosynostosis associated with microdeletion of chromosome 19p13.12-19p13.2. Genes Dis 2:347-352
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Shenaq, Deana S; Teven, Chad M; Seitz, Iris A et al. (2015) Characterization of Reversibly Immortalized Calvarial Mesenchymal Progenitor Cells. J Craniofac Surg 26:1207-13
Teven, Chad M; Farina, Evan M; Rivas, Jane et al. (2014) Fibroblast growth factor (FGF) signaling in development and skeletal diseases. Genes Dis 1:199-213
Beederman, Maureen; Farina, Evan M; Reid, Russell R (2014) Molecular basis of cranial suture biology and disease: Osteoblastic and osteoclastic perspectives. Genes Dis 1:120-125
Greives, Matthew R; Odessey, Eric A; Waggoner, Darrel J et al. (2013) RUNX2 quadruplication: additional evidence toward a new form of syndromic craniosynostosis. J Craniofac Surg 24:126-9
Chen, Liang; Jiang, Wei; Huang, Jiayi et al. (2010) Insulin-like growth factor 2 (IGF-2) potentiates BMP-9-induced osteogenic differentiation and bone formation. J Bone Miner Res 25:2447-59

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