HLS13-04 High-Content High-Throughput Screen for Diamond Blackfan Anemia Treatments

 描述(申请人提供)：有数千种罕见的遗传病没有得到批准的治疗方法。递归制药公司开发了一个药物发现平台，寻求将已知药物重新用于治疗此类疾病。该平台包括高含量免疫荧光图像分析，使用机器学习算法来识别RNAi和各种化学物质引起的相关和靶标变化。该系统已被用于在多种人类细胞类型中鉴定两个相关基因RPS19和RPS10功能丧失的表型。这些基因的突变是钻石黑粉贫血的主要原因，钻石黑粉贫血是一种罕见的遗传病，没有特定的靶向治疗。在这笔拨款中，我们建议：利用CRISPR/Cas9技术开发RPS19和RPS10基因敲除细胞株；利用我们的药物发现平台评估基因敲除细胞株的表型(‘Phenoprints’)；对数千种已知候选药物进行化学抑制物筛选，以确定哪些可以改善与RPS19和RPS10缺失相关的靶向Poprins；扩展我们的药物筛选平台的功能，以在我们的工作流程中使用非贴壁细胞系，并利用扩展的功能来确认药物识别为CD34+悬浮细胞培养模型中Blackfan贫血候选疗法的有效性。递归制药公司拥有执行这一第一阶段SBIR提案的经验、工具和动力，并加快该项目产生的任何化合物的商业开发。拟议的研究将具有重大的社会和商业影响。

 DESCRIPTION (provided by applicant): There are thousands of rare genetic diseases that have no approved treatment. Recursion Pharmaceuticals has developed a drug discovery platform that seeks to re-purpose known drugs for the treatment of such diseases. The platform consists of high content immunofluorescent image analysis using machine-learning algorithms to identify relevant and on-target changes induced by both RNAi and various chemicals. This system has been used to identify a phenotype for loss of function of two related genes, RPS19 and RPS10 in multiple human cell types. Mutations in these genes are the primary cause of Diamond Blackfan Anemia, a rare genetic disease with no specific targeted therapy. In this grant, we propose to: develop RPS19 and RPS10 knockout cell lines using CRISPR/Cas9 technology; evaluate knockout cell lines for phenotypes ('phenoprints') using our drug discovery platform; conduct chemical suppressor screens of thousands of known drug candidates to identify those that ameliorate on-target phenoprints associated with loss of RPS19 and RPS10; expand the capabilities of our drug screening platform to enable the use of non-adherent cell lines in our workflow and use the expanded capabilities to confirm the validity of drugs identifie as candidate therapies in CD34+ suspended cell culture models of Diamond Blackfan Anemia. Recursion Pharmaceuticals has the experience, tools, and drive to execute this Phase I SBIR proposal, and to accelerate commercial development of any compounds arising from the project. The proposed study would have significant societal and commercial implications.