This proposal investigates the use of induced pluripotent stem cell (iPSC)-derived endothelial cells (ECs) as surrogates for native ECs to improve our understanding and to better treat a life-threatening vascular disorder, pulmonary hypertension (PAH). To this end we have assembled a multidisciplinary team with expertise in clinical manifestations and pathobiology of PAH, iPSC generation and differentiation, genetics and genomics, bioinformatics and gene therapy and high throughput phosphoflow and microfluidic technologies. Our proposal reflects a unique opportunity to assess three lines of ECs derived from the same patient. This streamlines our ability to test the efficacy of iPSC-ECs as surrogates for native ECs, and to establish the significance of specific genetic alterations. We have a unique opportunity to compare gene variants, epigenetic and gene expression profiles in fibroblast derived iPSC-ECs to those in pulmonary arterial (PA)EC derived iPSC-ECs from the same PAH patients or controls. This should address in an unprecedented manner, how a genetic vulnerability leads to manifest disease. Moreover, we are well placed to investigate the application of gene and pharmaceutical therapy to reverse PAH pathology in iPSC-ECs. Because studies completed to date in the Wu laboratory have established facility with the production of iPSC-ECs, we are positioned to begin with Phase II of this Proposal. To this end, we propose three Specific Aims in Phase II and three in Phase III. Phase II, Aim 1 applies Hi-Seq, Methyl-Seq and RNA-Seq to obtain information on rare gene variants, epigenetic changes and gene expression in PAH and control fibroblast- iPSC-ECs, PAEC-iPSC-ECs and PAECs. Phase II, Aim 2 correlates this genomic information with function, by extensive analysis of the phenotype of these cells using angiogenesis assays and phosphoflow analyses and by assessing homogeneity using a single cell microfluidic approach. Phase II, Aim 3 initiates a partnership with Progenitor Cell Therapy to transfer the technology of producing iPSC-ECs so that it can be ramped up in the future to benefit large populations of patients. In Phase III, Aim 1 utilizes cutting edge gene therapy approaches, such as minicircles and ribosomal DNA vectors, to correct a gene variant in an iPSC-EC. Phase III, Aim 2 investigates how this reverts the PAH-related phenotype of the cell. Phase III, Aim 3 compares the efficacy of fibroblast and PAEC-iPSC-ECs vs. native PAECs to respond to pharmaceuticals to revert the disease phenotype. In summary, our studies should serve as a model to better understand how iPSC-ECs can be used to uncover and treat genetic predisposition to vascular dysfunction in a wide variety of cardiovascular diseases.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project--Cooperative Agreements (U01)
Project #
3U01HL107393-04S1
Application #
9062144
Study Section
Special Emphasis Panel (ZHL1-CSR-N (F1))
Program Officer
Gan, Weiniu
Project Start
2011-07-05
Project End
2016-06-30
Budget Start
2015-06-12
Budget End
2015-08-31
Support Year
4
Fiscal Year
2015
Total Cost
$8,140
Indirect Cost
$340
Name
Stanford University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94304
Zamanian, Roham T; Hedlin, Haley; Greuenwald, Paul et al. (2018) Features and Outcomes of Methamphetamine-associated Pulmonary Arterial Hypertension. Am J Respir Crit Care Med 197:788-800
Gu, Mingxia; Shao, Ning-Yi; Sa, Silin et al. (2017) Patient-Specific iPSC-Derived Endothelial Cells Uncover Pathways that Protect against Pulmonary Hypertension in BMPR2 Mutation Carriers. Cell Stem Cell 20:490-504.e5
Bonnet, SĂ©bastien; Provencher, Steeve; Guignabert, Christophe et al. (2017) Translating Research into Improved Patient Care in Pulmonary Arterial Hypertension. Am J Respir Crit Care Med 195:583-595
Saito, Toshie; Miyagawa, Kazuya; Chen, Shih-Yu et al. (2017) Upregulation of Human Endogenous Retrovirus-K Is Linked to Immunity and Inflammation in Pulmonary Arterial Hypertension. Circulation 136:1920-1935
Newman, John H; Rich, Stuart; Abman, Steven H et al. (2017) Enhancing Insights into Pulmonary Vascular Disease through a Precision Medicine Approach. A Joint NHLBI-Cardiovascular Medical Research and Education Fund Workshop Report. Am J Respir Crit Care Med 195:1661-1670
Yu, Kun-Hsing; Hart, Steven N; Goldfeder, Rachel et al. (2017) HARNESSING BIG DATA FOR PRECISION MEDICINE: INFRASTRUCTURES AND APPLICATIONS. Pac Symp Biocomput 22:635-639
Sa, Silin; Gu, Mingxia; Chappell, James et al. (2017) Induced Pluripotent Stem Cell Model of Pulmonary Arterial Hypertension Reveals Novel Gene Expression and Patient Specificity. Am J Respir Crit Care Med 195:930-941
Zhao, Ming-Tao; Shao, Ning-Yi; Hu, Shijun et al. (2017) Cell Type-Specific Chromatin Signatures Underline Regulatory DNA Elements in Human Induced Pluripotent Stem Cells and Somatic Cells. Circ Res 121:1237-1250
Matsa, Elena; Burridge, Paul W; Yu, Kun-Hsing et al. (2016) Transcriptome Profiling of Patient-Specific Human iPSC-Cardiomyocytes Predicts Individual Drug Safety and Efficacy Responses In Vitro. Cell Stem Cell 19:311-25
Hu, Shijun; Zhao, Ming-Tao; Jahanbani, Fereshteh et al. (2016) Effects of cellular origin on differentiation of human induced pluripotent stem cell-derived endothelial cells. JCI Insight 1:

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