Understanding and treating genetic urinary tract disease

先天性泌尿道疾病是个别罕见的，但在儿童和年轻人终末期肾病的主要原因。Urofacial syndrome（UFS）是一种常染色体隐性遗传疾病，可导致终生排尿障碍和潜在的致命性肾脏并发症，我们在UFS家族中发现了HPSE 2或LRIG 2突变，并描述了具有排尿缺陷和膀胱神经异常模式的Hpse 2和Lrig 2突变小鼠。在健康状态下，HPSE 2和LRIG 2在供应膀胱的神经中表达。因此，UFS膀胱疾病涉及周围神经病变。学生将加入我们的研究团队，并进行四个实验，如下，这将显著补充我们的工作计划，该计划部分由MRC项目资助MR/T016809/1遗传性膀胱疾病临床前基因治疗和英国肾脏研究项目资助Paed_RP_005_20190925先天性膀胱疾病的病理生理学。1.我们正在使用腺相关病毒（AAV）载体介导的HPSE 2或LRIG 2基因转移给婴儿小鼠，旨在改善Hpse 2或Lrig 2突变体的排尿缺陷。我们发现，未经治疗的UFS小鼠有受损的一氧化氮药物的膀胱流出的放松，以及膀胱体的过度收缩反应乙酰胆碱。学生将确定基因治疗是否与这些生理参数的改善有关。学生还将确定药理活性药物是否可以改善离体生理缺陷。这一点以及第1节（上文）将为更好的治疗铺平道路。学生将对未治疗和AAV治疗的UFS小鼠的肾脏进行组织学分析，以确定膀胱靶向基因治疗或药物治疗是否与肾脏和下肾道炎症的变化相关。学生将探讨产前基因转移的可能性，如报告基因表达评估，进入分化的肾脏道在体外，使用胚胎器官培养。这些使用AAV和慢病毒载体的“原理证明”实验将为产前治疗方案铺平道路。这项研究将为UFS或其他遗传性泌尿道疾病患者的基因治疗设计提供信息，就像使用AAV载体介导的脊髓性肌萎缩症基因治疗一样。

Congenital urinary tract diseases are individually rare but are collectively the main cause of end-stage renal disease in children and young adults. Urofacial syndrome (UFS) is one such disease, an autosomal recessive disorder causing lifelong impaired urinary voiding and potentially fatal kidney complications.We discovered HPSE2 or LRIG2 mutations in families with UFS and described Hpse2 and Lrig2 mutant mice with urination defects and abnormal patterns of bladder nerves. In health, HPSE2 and LRIG2 are expressed in nerves supplying the bladder. Thus, UFS bladder disease involves a peripheral neuropathy.The student will join our research team and undertake four experiments, below, that will markedly complement our programme of work that is in part funded by an MRC Project Grant MR/T016809/1 Preclinical gene therapy for genetic urinary bladder disease and a Kidney Research UK Project Grant Paed_RP_005_20190925 The pathophysiology of a congenital bladder disease.1. We are using adeno-associated virus (AAV) vector-mediated HPSE2 or LRIG2 gene transfer administered to baby mice aimed at ameliorating voiding defects in Hpse2 or Lrig2 mutants. We discovered that untreated UFS mice have impaired nitric-oxide medicated relaxation of the bladder outflow, as well as hypercontractility of the bladder body in response to acetylcholine. The student will determine whether gene therapy is associated with improvement in these physiological parameters.2. The student will also determine whether pharmacologically-active drugs can ameliorate the physiological defects ex vivo. This, as well as section 1 (above), will pave the way for better treatments.3. The student will perform histological analyses of kidneys of untreated and AAV-treated UFS mice to determine whether bladder-targeted gene therapy or pharmacological treatments are associated with changes in inflammation in the kidney and lower renal tract.4. The student will explore the possibility of prenatal gene transfer, as assessed by reporter gene expression, into the differentiating renal tract in vitro, using embryonic organ cultures. These 'proof of principle' experiments using AAV and lentivirus vectors will pave the way prenatal treatment protocols.This study will inform the design of gene therapy for people with UFS or other genetic urinary tract diseases, as being undertaken with AAV vector -mediated gene therapy for spinal muscular atrophy.