Renal Cell Engineering for Disease Modeling and in vivo Regeneration

• Applicant: Dr. Michael Kaminski
• Subject Area: Nephrology
• Term: since 2019
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 418458827

Forced expression of transcription factors can change the fate of somatic cells, directing them to alternate cell fates in a process called direct reprogramming or transdifferentiation. This technique represents an easy and fast approach for the generation of desired cell types in vitro and in vivo. One practical use of direct reprogramming is in vitro disease modeling. Here, a cellular model, which reflects basic properties of the disease, allows for investigation of the disease-specific phenotype, pathogenesis and potential therapeutic interventions. The derivation of cell sources for reprogramming from affected patients allows for investigation of the disease on a patient-specific genetic background. Further, overexpression of transcription factors in vivo enables regeneration of tissue lost through injury or disease. This strategy has been proven to be successful in mouse models of myocardial infarction, diabetes and liver fibrosis.In my previous work, we succeeded in generating renal tubular epithelial cells (iRECs) from human and mouse fibroblasts through the overexpression of 4 transcription factors. The resulting cells resembled their native counterparts in morphology, transcriptome, metabolome and function. The main goal of the proposed research is to improve renal reprogramming in order to generate nephron segment-specific renal cell types and to use renal reprogramming for disease modelling and in vivo regeneration. The results will contribute to a better understanding of the transcriptional control of renal cell identity and help deciphering the molecular causes of some forms of genetic kidney disease. Further, renal in vivo reprogramming will explore new avenues for regenerative medicine applications. To reach this overall goal, we will apply novel synthetic biology tools, which we have developed at MIT. This includes genetic reporters of renal cell fate, enabling high throughput screening of candidate reprogramming factors, as well as synthetic genetic feedback circuits to precisely steer the expression level of reprogramming factors. Building on our previous work on congenital anomalies of the kidney and urinary tract (CAKUT), we will employ direct reprogramming to derive renal cells from skin cells of affected patients and investigate CAKUT pathogenesis in a patient-specific manner. Finally, we will use mouse models of fibrosis and ischemia reperfusion injury to test the potential of direct reprogramming to regenerate tissue function in vivo. With this project, we aim to contribute to our understanding of the molecular mechanisms of kidney diseases and establish a novel regenerative approach that may be translated into future clinical practice.

• DFG Programme: Independent Junior Research Groups