Gibson Christopher C, Zhu Weiquan, Davis Chadwick T, Bowman-Kirigin Jay A, Chan Aubrey C, Ling Jing, Walker Ashley E, Goitre Luca, Delle Monache Simona, Retta Saverio Francesco, Shiu Yan-Ting E, Grossmann Allie H, Thomas Kirk R, Donato Anthony J, Lesniewski Lisa A, Whitehead Kevin J, Li Dean Y
From the Program in Molecular Medicine (C.C.G., W.Z., C.T.D., J.A.B.-K., A.C.C., J.L., A.H.G., K.R.T., K.J.W., D.Y.L.), Department of Bioengineering (C.C.G., Y.-T.E.S.), Department of Medicine (C.C.G., W.Z., K.R.T., D.Y.L.), Department of Human Genetics (C.T.D.), Department of Oncological Sciences (A.C.C., D.Y.L.), Division of Geriatrics, Department of Medicine (A.E.W., A.J.D., L.A.L.), Division of Nephrology and Hypertension, Department of Medicine (Y.-T.E.S.), Department of Pathology (A.H.G.), Division of Cardiology, and Department of Medicine (K.J.W., D.Y.L.), University of Utah, Salt Lake City, UT; Recursion Pharmaceuticals, LLC, Salt Lake City, UT (C.C.G., D.Y.L.); CCM Italia, Department of Clinical and Biological Sciences, University of Torino, Orbassano, Torino, Italy (L.G., S.F.R.); CCM Italia, Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy (S.D.M.); Geriatrics Research Education and Clinical Center, Veteran's Affairs Medical Center, Salt Lake City, UT (A.J.D., L.A.L.); The Key Laboratory for Human Disease Gene Study of Sichuan Province, Institute of Laboratory Medicine, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu, Sichuan, China (D.Y.L.); and Cardiology Section, VA Salt Lake City Health Care System, Salt Lake City, UT (K.J.W., O.Y.L.).
Circulation. 2015 Jan 20;131(3):289-99. doi: 10.1161/CIRCULATIONAHA.114.010403. Epub 2014 Dec 8.
Cerebral cavernous malformation (CCM) is a hemorrhagic stroke disease affecting up to 0.5% of North Americans that has no approved nonsurgical treatment. A subset of patients have a hereditary form of the disease due primarily to loss-of-function mutations in KRIT1, CCM2, or PDCD10. We sought to identify known drugs that could be repurposed to treat CCM.
We developed an unbiased screening platform based on both cellular and animal models of loss of function of CCM2. Our discovery strategy consisted of 4 steps: an automated immunofluorescence and machine-learning-based primary screen of structural phenotypes in human endothelial cells deficient in CCM2, a secondary screen of functional changes in endothelial stability in these same cells, a rapid in vivo tertiary screen of dermal microvascular leak in mice lacking endothelial Ccm2, and finally a quaternary screen of CCM lesion burden in these same mice. We screened 2100 known drugs and bioactive compounds and identified 2 candidates, cholecalciferol (vitamin D3) and tempol (a scavenger of superoxide), for further study. Each drug decreased lesion burden in a mouse model of CCM vascular disease by ≈50%.
By identifying known drugs as potential therapeutics for CCM, we have decreased the time, cost, and risk of bringing treatments to patients. Each drug also prompts additional exploration of biomarkers of CCM disease. We further suggest that the structure-function screening platform presented here may be adapted and scaled to facilitate drug discovery for diverse loss-of-function genetic vascular disease.
脑海绵状血管畸形（CCM）是一种出血性中风疾病，影响着多达0.5%的北美人，目前尚无获批的非手术治疗方法。一部分患者患有该疾病的遗传形式，主要是由于KRIT1、CCM2或PDCD10基因的功能丧失突变。我们试图确定可以重新用于治疗CCM的已知药物。
我们基于CCM2功能丧失的细胞和动物模型开发了一个无偏倚的筛选平台。我们的发现策略包括4个步骤：对缺乏CCM2的人内皮细胞进行基于自动免疫荧光和机器学习的结构表型初步筛选，对这些相同细胞中内皮稳定性的功能变化进行二次筛选，对缺乏内皮Ccm2的小鼠进行快速的体内三级筛选以检测真皮微血管渗漏，最后对这些相同小鼠的CCM病变负担进行四级筛选。我们筛选了2100种已知药物和生物活性化合物，确定了2种候选药物，胆钙化醇（维生素D3）和tempol（一种超氧化物清除剂），用于进一步研究。每种药物使CCM血管疾病小鼠模型中的病变负担降低了约50%。
通过将已知药物确定为CCM的潜在治疗方法，我们减少了将治疗方法带给患者的时间、成本和风险。每种药物还促使对CCM疾病的生物标志物进行进一步探索。我们进一步建议，这里介绍的结构-功能筛选平台可以进行调整和扩展，以促进针对各种功能丧失性遗传性血管疾病的药物发现。
