iPS-technology and patient specific disease models

iPSC技术的开发和应用有望彻底改变我们对疾病机制的理解，并提高治疗方案的可用性。产生患者特异性iPSC的能力允许开发基于细胞的疾病建模，其中以前极难获得足够量的相关人类细胞类型以进行研究，例如心肌细胞、内皮细胞（EC）和血管平滑肌细胞（VSMC）。此外，该技术能够研究心血管疾病的潜在机制。 迄今为止，我们已经成功地生成了一个人类iPSC生物库，其中包含来自健康志愿者和携带ACDC、HIES（乔布斯综合征）、SAVI、CADASIL、DADA 2、奥拉斯、特纳综合征和Degos等遗传缺陷的患者的几个样本。大多数iPSC系也被广泛表征为表现出无限自我更新的潜力以及分化成三个胚层的能力。此外，我们已经建立了新的分化方案，其具有有效的、分阶段的、化学定义的策略，用于中胚层谱系细胞（EC、MSC、VSMC、造血谱系细胞以及使用这些细胞形成血管类器官）的短期诱导。这种方法已被证明在增加我们对疾病病理生理学的理解以及允许测试药物/小分子作为潜在的新型治疗策略或细胞替代疗法方面至关重要。 使用这些技术与ACDC，我们已经证明了TNAP的补偿性上调，导致腺苷的产生不足和PPi的显着减少，导致体外ACDC样品中的血管钙化。在体内，我们还确定了A2 B腺苷受体信号传导途径下游的抑制途径和几种治疗药物靶点，其中依替膦酸盐是临床上最可行的候选药物。另一方面，当关注由信号转导子和转录激活子3（STAT 3）的功能丧失突变引起的AD-HIES的潜在机制时，我们能够确定STAT 3通过Nanog的激活在体细胞向iPSC的重编程中起关键作用。我们还开发了一个畸胎瘤模型（小鼠）系统与iPSC，以探索治疗AD-HIES的药理学靶点。这些iPSC细胞系及其衍生物完成的研究在过去一年中在《自然》、《干细胞研究》和《毒理学化学研究》上发表了大量论文。 我们计划继续应用这些策略来研究遗传原因和疾病机制，并对DADA 2，奥拉斯，Turner综合征和Degos以及其他可能具有不清楚遗传原因或病理机制的血管表型的罕见疾病进行持续研究。

The development and application of iPSC technology promises to revolutionize our understanding of disease mechanisms and improve the availability of treatment options. The ability to generate patient specific iPSC allows for the development of cell-based disease modeling where it has previously been extremely difficult to obtain sufficient amounts of the relevant human cell types to study such as cardiomyocytes, endothelial (ECs) and vascular smooth muscle cells (VSMCs). Further, this technology enables the investigation of mechanisms underlying cardiovascular diseases. We have to-date successfully generated a human iPSC biobank with several samples derived from healthy volunteers and patients carrying genetic defects for conditions such as ACDC, HIES (Jobs syndrome), SAVI, CADASIL, DADA2, ORAS, Turner Syndrome and Degos. The majority of iPSC lines have also been extensively characterized to exhibit the potential of unlimited self-renewal as well as the ability to differentiate into three germ layers. Additionally, we have established novel differentiation protocols with efficient, stage-wise, chemically-defined strategies for short-term induction of mesoderm lineage cells (ECs, MSCs, VSMCs, Hematopoietic lineage cells as well as the formation of blood vessel organoids using these cells). This approach has proven critical in increasing our understanding of the disease pathophysiology as well as allowed for testing of drugs/small molecules as potential novel therapeutic strategies or cell replacement therapy. Using these technologies with ACDC, we have demonstrated a compensatory upregulation of TNAP leading to an insufficient production of adenosine and a marked decrease in PPi, causing the vascular calcifications in ACDC samples from patients in vitro. In vivo, we also identified an inhibitory pathway downstream of the A2B adenosine receptor signaling pathway and several therapeutic drug targets, with etidronate being the most clinically viable candidate. On the other hand, when focusing on the underlying mechanisms of AD-HIES that is caused by loss-of-function mutations in signal transducer and activator of transcription 3 (STAT3), we were able to identify that STAT3 plays a critical role in reprogramming of somatic cells towards iPSCs through activation of Nanog. We alsodeveloped a teratoma model (murine) system with iPSC to explore pharmacological targets for treatment of AD-HIES. The research accomplished with these iPSC lines and their derivatives has led to a large volume of publications in Nature, Stem Cell Research, and Chemical Research in Toxicology over the last year. We plan to continue to apply these strategies to study the genetic cause and disease mechanisms with on-going research for DADA2, ORAS, Turner Syndrome and Degos as well as for other rare conditions with vascular phenotype that may have unclear genetic cause or pathomechanisms.