干细胞生物学报告
题目：Role of innate immunity in nuclear reprogramming of cell fate
主讲：John P Cooke
Houston Methodist Research Institute, Cardiovascular Sciences, Houston, TX, 77030
时间：2013年10月22日（周二）上午10:00-11:00 AM
地点：生命科学学院311会议室
联系人：邓宏魁（电话：62756474）
Background: Cell-based approaches to regenerate the endothelium holds promise, with one such candidate source being induced pluripotent stem cells (iPSCs). We have investigated the potential of endothelial cells (ECs) derived from iPSCs to promote the perfusion of ischemic tissue in a murine model of peripheral artery disease (PAD).However, it may be more efficient to transdifferentiate fibroblasts to ECs directly. Recent reports have suggested that direct reprogramming to ECs is feasible, however still requires the use of viral vectors encoding transcription factors, thus making them clinically unsafe.
Hypothesis: We recently discovered that retroviral vectors encoding the reprogramming factors, by activating the Toll-like receptor 3 (TLR3) pathway, make nuclear reprogramming possible by increasing epigenetic plasticity and favoring an open chromatin state (Lee & Sayed et al. Cell). Based on this recognition that innate immunity favors an open chromatin state, we hypothesized that activation of TLR3, together with external microenvironmental cues that drive EC specification, might induce transdifferentiation of fibroblasts into ECs (“iECs”).
Results: Intriguingly, we find that TLR3 agonist Poly I:C, combined with exogenous endothelial growth factors, is sufficient to transdifferentiate fibroblasts into iECs (in the absence of viral vectors or transcription factors). These iECs exhibited all the characteristics of EC phenotype comparable to HMVECs including ability to form capillarylike structures and incorporating acetylated-LDL. Furthermore, iECs significantly improved limb perfusion and neovascularization in the ischemic hindlimb. RNAseq studies indicate high concordance of iEC and EC gene expression. Finally, loss-of function studies reveal that innate immune pathways are critical for efficient transdifferentiation.
Conclusion: This study is a first step toward development of a regenerative strategy using ECs derived from small molecules and growth factors without use of viral vectors encoding transcription factors. Moreover, we find that innate immune pathways are critical for efficient transdifferentiation.
Selected Publications:
1. Jang J, Ho H-K, Kwan HH, Adimoolam S, Fajardo LF, Cooke JP: Angiogenesis is impaired by hypercholesterolemia: role of asymmetric dimethylarginine. Circulation 2000 Sep 19;102(12):1414-9.
2. Uemura S, Fathman G, Rothbard J, Cooke JP: Rapid and efficient vascular transport of arginine polymers inhibits myointimal hyperplasia. Circulation 2000 Nov 21;102(21):2629-35.
3. Heeschen C, Jang J, Hoai-Ky V, Kaji S, Yang P, Hu RS, Cooke JP: Nicotine is an agent of angiogenesis. Nicotine stimulates angiogenesis and promotes tumor growth and atherosclerosis. Nat Med 2001 Jul; 7(7):833-9.
4. Lin KY, Ito A, Asagami T, Tsao PS, Adimoolam S, Kimoto M, Tsuji H, Reaven G, Cooke JP: Impaired nitric oxide synthase pathway in diabetes mellitus: Role of asymmetric dimethylarginine and dimethylarginine dimethylaminohydrolase. Circulation 2002 Aug 20;106(8):987-92.
5. Heeschen C, Weis M, Cooke JP: A novel angiogenic pathway mediated by non-neuronal nicotinic acetylcholine receptors. J Clin Invest 2002 Aug;110(4):527-36.
6. Aicher A, Heeschen C, Mohaupt M, Cooke JP, Zeiher AM , and Dimmeler S: Nicotine Strongly Activates Dendritic Cell-Mediated Adaptive Immunity – Potential Role for Progression of Atherosclerotic Lesions. Circulation. 2003 Feb 4;107(4):604-11.
7. Zhu B, Heeschen C, Sievers RE, Karliner JS, Parmley WW, Glantz SA and Cooke JP: Second Hand Smoke Stimulates Tumor Angiogenesis and Growth, Cancer Cell 2003; 4(3):191-6.
8. Dayoub H, Achan V, Adimoolam S, Jacobi J, Stuehlinger M, Wang B, Tsao PS, Kimoto M, Vallance P, Patterson AJ, Cooke JP: DDAH Regulates NO Synthesis: Genetic and physiological evidence. Circulation 2003; 108: 1043-1048.
9. Weis M, Kledal TN, Lin KY, Panchal SN, Gao SZ, Valantine HA, Mocarski ES, Cooke JP: Cytomegalovirus infection impairs the NOS pathway. Role of ADMA in transplant arteriosclerosis. Circulation 2004 Feb 3;109(4):500-5.
10. Fearon WF, Aarnoudse W, Pijls NH, De Bruyne B, Balsam LB, Cooke DT, Robbins RC, Fitzgerald PJ, Yeung AC, Yock PG. Microvascular resistance is not influenced by epicardial coronary artery stenosis severity: experimental validation. Circulation 2004; 109(19):2269-2272.
11. Jacobi J, Sydow K, von Degenfeld G, Zhang Y, Dayoub H, Wang B, Patterson AJ, Kimoto M, Blau HM, Cooke JP: Overexpression of Dimethylarginine Dimethylaminohydrolase (DDAH) Reduces Tissue ADMA Levels and Enhances Angiogenesis. Circulation 2005 Mar 22;111(11):1431-8.
12. Tu W, Potena L, Stepick-Biek P, Liu L, Dionis KY, Luikart H, Fearon WF, Holmes TH, Cooke JP, Valantine HA, Mocarski ES and Lewis DB: T-Cell immunity to subclinical cytomegalovirus infection reduces cardiac allograft disease. Circulation 2006;114:1608-1615.
13. Wilson A, Harada R, Nair N, Balasubramanian N, Cooke JP: L-arginine supplementation in peripheral arterial disease: No benefit and possible harm. Circulation 2007 Jul 10;116(2):188-95.
14. Li Z, Wu JC, Sheikh AY, Kraft D, Cao F-, Xie X, Robbins RC, Cooke JP, Wu JC: Differentiation, survival, and function of embryonic stem cell-derived endothelial cells for ischemic heart disease.Circulation 2007 Sep 11;116(11 Suppl):I46-54.
15. Wilson AM, Kimura E, Harada RK, Nair N, Narasimhan B, Meng X-Y, Zhang F, Beck KR, Olin JW, Fung ET, Cooke JP: Beta-2 microglobulin as a biomarker in peripheral arterial disease. Proteomic profiling and clinical studies. Circulation. 2007 Sep 18;116(12):1396-403.
16. Wu JC, Chruscinski A, de Jesus Perez VA, Singh H, Pitsiouni M, Rabinovitch M, Utz PJ, Cooke JP : Cholinergic modulation of angiogenesis : Role of the α7 nicotinic acetylcholine receptor. J Cell Biochem 2009, Oct 1;108(2):433-46.
17. Holweg CTJ, Potena L, Luikart H, Yu T, Berry GJ, Cooke JP, Valantine HA, Mocarski ES: Identification and classification of acute cardiac rejection by intragraft transcriptional profiling.Circulation. 2011 May 24;123(20):2236-43.
18. Hong G, Lee JC, Robinson JT, Raaz U, Xie L, Huang NF, Cooke JP and Dai H: Multi-functional in vivo vascular imaging using near-infrared II fluorescence. Nat Med. 2012 Dec 6;18(12):1841-6.
19. Lee J, Sayed N, Hunter A, Au KF, Wong WH, Mocarski E, Reijo Pera R, Cooke JP: Activation of innate immunity is required for efficient nuclear reprogramming. Cell. 2012 Oct 26;151(3):547-58.
20. Tian XY, Wong WT, Wang N, Lu Y, Cheang WS, Liu Y, Lee SS, Chen Z-Y, Yao X, Cooke JP, Huang Y: PPAR δ activation protects endothelial function in diabetic mice. Diabetes. 2012 Dec;61(12):3285-93.
21. Ahn G-O, Seita J, Lee J, Leeper NJ, Cooke JP, Weissman IL, Brown JM: Activation of hypoxia- inducible factor-1 (HIF-1) in myeloid cells is sufficient to mediate angiogenic effects in mice PNAS May 9, 2012
欢迎各位老师和同学积极参加！