Modulation of cyst growth in renal tissue from patients with autosomal dominant polycystic kidney disease in a human tissue-based 3D-in-vivo-model

常染色体显性遗传性多囊肾病（ADPKD）患者双侧多发性肾囊肿逐渐增大，导致肾功能下降。细胞增殖和液体分泌是囊肿扩大的两个基本特征。目前，仍然需要抑制囊肿生长的疗法，以保护患有ADPKD的患者的肾功能。我们小组的最新数据表明，ADPKD小鼠模型中的囊肿生长可通过TMEM 16 A（Anoctamin 1）介导的三种不同的跨上皮氯分泌药理学抑制剂来预防。此外，我们在小鼠模型中发现缺氧和随后的缺氧诱导因子HIF-1α的激活促进了囊肿的生长。接下来，我们的目标是将这些发现转化为临床应用和我们的患者。作为一个重要的中间步骤，我们需要测试最有希望的药物在人体组织中的疗效。我们在人类肿瘤研究领域拥有长期使用3D体内CAM（绒毛尿囊膜）模型的专业知识，并建立了CAM模型来研究不同疗法（化疗，放疗）对肿瘤生长和血管生成的影响。除了肿瘤组织，我们还成功地培养了人类ADPKD肾组织一周多，我们能够在各种条件下监测囊肿生长。该模型应该使我们-除了动物-和细胞培养模型-回答以下问题：1。氯化物分泌抑制剂（例如TMEM 16 A抑制剂，不同浓度）对CAM上人类多囊肾组织囊肿生长的影响是什么？肾脏的不同部位或ADPKD患者对某些药物的反应是否不同？2.在CAM模型中，批准的脯氨酰羟化酶抑制剂对囊肿生长有什么影响？3.囊肿液成分对囊肿扩大有影响吗？哪些因素会影响流体成分，以及如何改变流体成分？4.是否有可能在CAM上培养小鼠肾脏切片来研究肾脏（病理）生理学，从而提供除多囊肾疾病之外的大量肾脏相关分析？通过这项工作，我们的目标是深入了解ADPKD患者对不同剂量的几种药物的反应。在这个过程中，我们希望提高我们对ADPKD病理机制的理解，并确定新的药理学靶点。 总之，新方法是开发和测试创新治疗方法的重要一步，随后有助于根据3R原则（“减少，替换，改进”）减少动物实验。

In autosomal dominant polycystic kidney disease (ADPKD) multiple bilateral renal cysts gradually enlarge, leading to a decline in renal function. Cell proliferation and fluid secretion are two essential characteristics of cyst enlargement. Currently, there is still a need for therapies that inhibit cyst growth in order to preserve renal function in patients suffering from ADPKD. Recent data from our group has shown that cyst growth in an ADPKD mouse model is prevented by three different pharmacological inhibitors of transepithelial chloride secretion mediated by TMEM16A (Anoctamin 1). Furthermore, we have shown that hypoxia and subsequent activation of the hypoxia-inducible factor HIF-1α promotes cyst growth in our mouse model.Next, we aim to translate these findings into clinical application and our patients. As an important intermediate step, we need to test the most promising drugs with regard to their efficacy in human tissue. We have long-standing expertise using the 3D in vivo CAM (chorioallantoic membrane) model in the area of human tumor research and have established the CAM model to study the effects of different therapies (chemotherapy, radiation therapy) on tumor growth and angiogenesis. In addition to tumor tissue, we were successful in cultivating human ADPKD kidney tissue for more than a week and we were able to monitor cyst growth under various conditions. The model should enable us – in addition to animal- and cell culture models – to answer the following questions:1. What are the effects of inhibitors of chloride secretion (e.g. TMEM16A inhibitors, different concentrations) on cyst growth of human polycystic kidney tissue on the CAM? Do different parts of the kidney or individual ADPKD patients respond differently to certain drugs?2. What are the consequences of approved prolyl-hydroxylase inhibitors on cyst growth in the CAM model?3. Does cyst fluid composition have an impact on cyst enlargement? Which factors influence fluid composition and how can it be changed pharmacologically?4. Is it possible to culture mouse kidney slices on the CAM to study kidney (patho-) physiology which would offer a plethora of kidney-related analyses beyond polycystic kidney disease? With the proposed work, we aim to gain insights into the response of individual ADPKD patients to several drugs with different dosages. During the course of this process we hope to improve our understanding of the pathological mechanisms involved in ADPKD and identify novel pharmacological targets. In summary, the new methodology represents an important step in the development and testing of innovative therapeutic approaches and subsequently contributes to the reduction of animal experiments according to the 3R-principles („reduce, replace, refine“).