Personalised medicine in Joubert syndrome: patient-specific human urine-derived epithelial cells as a disease model and platform for therapeutic devel

在英国需要透析/移植的4万名患者中，“纤毛病”占10%。这些疾病导致囊性肾病、视网膜变性和脑异常，如Joubert综合征（JS），但广泛的基因型-表型异质性对解剖疾病病理生物学提出了挑战。目前还没有针对这些疾病的改善治疗方法，治疗方法的发展将需要针对患者的个性化方法。导师（https://www.ncl.ac.uk/igm/staff/profile/johnsayer.html#background和https://www.ncl.ac.uk/igm/staff/profile/colinmiles.html#background）已经从基因发现中开发了一个“管道”[例如，Nat Genet. 2006 38(6):674；[J] .中国生物医学工程学报，2018,35(4):612。人类尿源性肾上皮细胞（human Urine-derived-Renal-Epithelial-Cells, hURECs） [j] .中华医学杂志，2017,26(23):4657；中华医学杂志，2018][https://www.chroniclelive.co.uk/news/health/newcastle-university-kidney-research-sayer-15431046], 2014 111(27):9893]。虽然主管们在涉及hURECs的研究领域处于领先地位，但这些细胞仍然是该管道中最不为人所知的部分，进一步详细了解和开发该平台对于将我们的发现转化为临床至关重要。目前，涉及hURECs的研究仅限于实验室几个小时内的患者，迫切需要克服这一限制，例如，我们在世界各地（从美国到中东）都有患者，但目前必须将我们的研究人员派往患者所在的地方。因此，与此同时，该项目将与专门储存和运输人类细胞的先驱Atelerix （https://www.atelerix.co.uk/）合作进行。该项目将包括对JS患者的hURECs进行详细的表型表征，并将其与健康志愿者的hURECs和已被设计（siRNA和/或CRISPR/Cas9）以模拟疾病特定方面的hURECs进行比较。除了标准的免疫组织化学之外，该项目的关键使能技术是基因组学核心设施，该设施得到了纽卡斯尔大学最近100万英镑的投资（https://www.ncl.ac.uk/igm/research/facilities/genomicscf/）的支持，这将使我们能够在hURECs上执行“单细胞RNAseq”，以前所未有的细节表征这些细胞。这些数据将被叠加在正常肾脏数据集上，以精确地定义hUREC培养物的细胞组成，随后，疾病机制将通过与患者特异性小鼠模型（即，经过改造的小鼠携带与患者相同的氨基酸变化）的数据进行比较来确定。该博士学位将提供最先进的人类疾病遗传学的综合培训，并结合工业角度的基本技术的发展，由一个多学科团队在纤毛病研究的前沿提供。

"Ciliopathies" account for 10% of the 40,000 UK patients requiring dialysis/transplantation. These disorders cause cystic kidney disease, retinal degeneration and brain abnormalities, as typified by Joubert syndrome (JS) but widespread genotype-phenotype heterogeneity poses a challenge for dissecting disease pathobiology. There are currently no disease-modifying treatments for these conditions and the development of treatments will require patient-specific, personalised approaches. The supervisors (https://www.ncl.ac.uk/igm/staff/profile/johnsayer.html#background and https://www.ncl.ac.uk/igm/staff/profile/colinmiles.html#background) have developed a "pipeline" from gene discovery [eg, Nat Genet. 2006 38(6):674; Am J Hum Genet. 2018 103(4):612], through creation of mouse models [eg, Proc Natl Acad Sci USA. 2014 111(27):9893] for identification of therapeutic targets, and back to the patient for testing and validation of potential treatments on kidney cells directly isolated from patients [eg, Hum Mol Genet. 2017 26(23):4657 & Proc Natl Acad Sci 2018]], known as human Urine-derived-Renal-Epithelial-Cells (hURECs)n https://www.chroniclelive.co.uk/news/health/newcastle-university-kidney-research-sayer-15431046. Whilst the supervisors lead the field in research involving hURECs, these cells remain the least understood part of this pipeline and further detailed understanding and development of this platform will be essential for translating our findings to the clinic. Currently studies involving hURECs are limited to those patients within a few hours of the laboratory and there is an urgent need to overcome this limitation, eg, we have patients worldwide (from USA to the Middle East) but at present must send our researchers to where the patients are. Therefore, in parallel, this project will be carried out in collaboration with Atelerix (https://www.atelerix.co.uk/) pioneers in the storage and transport of specialised human cells. The project will involve detailed phenotypic characterisation of hURECs from a panel of JS patients compared with hURECs from healthy volunteers and hURECs that have been engineered (siRNA and/or CRISPR/Cas9) to model specific aspects of disease. In addition to standard immunohistochemistry, a key enabling technology for this project is the Genomics Core Facility underpinned by recent >£1M investments from Newcastle University (https://www.ncl.ac.uk/igm/research/facilities/genomicscf/ ) that will allow us to perform "single cell RNAseq" on hURECs to characterise these cells in unprecedented detail. These data will be overlayed on normal kidney datasets to precisely define the cellular composition of hUREC cultures and subsequently, disease mechanisms will be identified by comparison with data from patient-specific mouse models (ie, mice engineered to carry the identical amino acid change as the patient). This PhD will provide a comprehensive training in state-of-the-art human disease genetics coupled with an industrial perspective on the development of essential technologies, delivered by a multidisciplinary team at the forefront of ciliopathy research.