The ability to generate induced pluripotent stem cells (iPSCs) from skin fibroblasts and other mature cells and then to drive them to distinct lineages brings closer the promise of cell based therapies. Here we propose to test the feasibility of using iPSC-derived endothelial progenitor cells (EPCs) alone or in conjunction with hematopoietic stem cells (HSCs) to induce restoration of lung endothelial barrier function and regeneration of lung vessels after lung injury induced by sepsis. We will also define the molecular and cellular mechanisms mediating endothelial barrier and vascular repair induced by the iPSC-derived progenitors. The possibility of using iPSC-derived progenitor cells represents a quantal advancement over current approaches employing the difficult to obtain and controversial ESC-derived cells or mesenchymal cells with consequences and short-lived humorally-mediated actions. However, there are fundamental questions concerning the use, safety, and mechanism of action of iPSC- derived cells, which this proposal hopes to address. These questions include do these progenitors cells have a """"""""memory"""""""" of their origin, do they function as effectively as ESC-derived progenitor cells when differentiated into EPCs, are their effects secondary to engraftment in the injured lung niche and/or humoral mechanisms, do they repair the injured endothelial barrier and lung vessels and if so how. The studies are based on our ability to differentiate iPSCs to a relatively pure population of EPCs (which are positive for both Flk1 and VE-cadherin markers) and are characterized by their ability to form an endothelial barrier and produce blood vessels. We propose the following aims:
Aim 1, To address the endothelial barrier protective and vascular regenerative potential of mouse iPSCs differentiated into EPCs and HSCs;
Aim 2, To determine the role of iPSC-derived EPCs in restoring lung microvascular barrier function and fluid balance in sepsis;
and Aim 3, To determine the role of iPSC-derived progenitor cells in mediating microvascular regeneration after lung vascular injury. These studies will be made using diverse approaches requiring the efforts and expertise of a team of outstanding multi-disciplinary investigators and consultants, with the hope of providing the scientific rationale for iPSC-based therapy targeted against acute inflammatory injury.

Public Health Relevance

Sepsis-induced acute lung injury (ALI) remains an intractable disease with a mortality of 50% or greater despite advanced interventions. ALI bears the hallmark of inflammatory infiltration and protein-rich edema fluid due to severe disruption of lung vascular endothelial barrier. Our studies will assess the effectiveness and mechanisms for that effectiveness of the transplantation of induced pluripotent stem cells (iPSC), derived from mouse fibroblasts, and subsequently differentiated into endothelial progenitor cells (EPCs) and hematopoietic stem cells (HSCs). These cells will be studied individually or in combination to see if they can reverse established lung microvessel injury that is induced by two sepsis mouse models and thereby re-establish normal lung fluid balance. We have generated these mouse progenitor and stem cells and found that we can readily expand them, a requirement for cell-based therapy. We hope by understanding the mechanisms of the protective effects of these easily obtainable progenitor cells to lay rigorous scientific underpinnings for the eventual use of defined iPSC-derived progenitor cells for the treatment of ALI in humans.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL090152-06
Application #
8282697
Study Section
Respiratory Integrative Biology and Translational Research Study Section (RIBT)
Program Officer
Blaisdell, Carol J
Project Start
2007-09-28
Project End
2016-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
6
Fiscal Year
2012
Total Cost
$481,820
Indirect Cost
$175,820
Name
University of Illinois at Chicago
Department
Pharmacology
Type
Schools of Medicine
DUNS #
098987217
City
Chicago
State
IL
Country
United States
Zip Code
60612
Gong, Haixia; Liu, Menglin; Klomp, Jeff et al. (2017) Method for Dual Viral Vector Mediated CRISPR-Cas9 Gene Disruption in Primary Human Endothelial Cells. Sci Rep 7:42127
Tsang, Kit Man; Hyun, James S; Cheng, Kwong Tai et al. (2017) Embryonic Stem Cell Differentiation to Functional Arterial Endothelial Cells through Sequential Activation of ETV2 and NOTCH1 Signaling by HIF1?. Stem Cell Reports 9:796-806
Marsboom, Glenn; Chen, Zhenlong; Yuan, Yang et al. (2017) Aberrant caveolin-1-mediated Smad signaling and proliferation identified by analysis of adenine 474 deletion mutation (c.474delA) in patient fibroblasts: a new perspective on the mechanism of pulmonary hypertension. Mol Biol Cell 28:1177-1185
Yamada, Kaori H; Kang, Hojin; Malik, Asrar B (2017) Antiangiogenic Therapeutic Potential of Peptides Derived from the Molecular Motor KIF13B that Transports VEGFR2 to Plasmalemma in Endothelial Cells. Am J Pathol 187:214-224
Zhang, Lianghui; Jambusaria, Ankit; Hong, Zhigang et al. (2017) SOX17 Regulates Conversion of Human Fibroblasts Into Endothelial Cells and Erythroblasts by Dedifferentiation Into CD34+ Progenitor Cells. Circulation 135:2505-2523
Cantelmo, Anna Rita; Conradi, Lena-Christin; Brajic, Aleksandra et al. (2016) Inhibition of the Glycolytic Activator PFKFB3 in Endothelium Induces Tumor Vessel Normalization, Impairs Metastasis, and Improves Chemotherapy. Cancer Cell 30:968-985
Klomp, Jennifer E; Huyot, Vincent; Ray, Anne-Marie et al. (2016) Mimicking transient activation of protein kinases in living cells. Proc Natl Acad Sci U S A 113:14976-14981
Gong, Haixia; An, Shejuan; Sassmann, Antonia et al. (2016) PAR1 Scaffolds TGF?RII to Downregulate TGF-? Signaling and Activate ESC Differentiation to Endothelial Cells. Stem Cell Reports 7:1050-1058
Marsboom, Glenn; Zhang, Guo-Fang; Pohl-Avila, Nicole et al. (2016) Glutamine Metabolism Regulates the Pluripotency Transcription Factor OCT4. Cell Rep 16:323-332
Wu, Liangtang; Wary, Kishore K; Revskoy, Sergei et al. (2015) Histone Demethylases KDM4A and KDM4C Regulate Differentiation of Embryonic Stem Cells to Endothelial Cells. Stem Cell Reports 5:10-21

Showing the most recent 10 out of 33 publications