Establishment of miR-21 gene editing by CRISPR/Cas9 in an in vitro model of metastatic clear cell renal cell Carcinoma

• Grant number: 22/02246-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2022
• End date: May 31, 2023
• Field of knowledge: Health Sciences - Medicine - Surgery
• Principal Investigator: Sabrina Thalita dos Reis Faria
• Grantee: Caroline Brandão Chiovatto
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Renal Cell Carcinoma (RCC) is a malignant, chemotherapy-resistant neoplasm that originates in the proximal contorted tubule lining of nephrons. The disease presents approximately 4% of malignancies in adults and was ranked in 2018 as the sixth deadliest cancer worldwide. RCC can be classified according to distinct genetic, molecular, and morphological features and is composed of different subtypes. Clear cell RCC is the most prevalent subtype and accounts for about 75% of all RCCs. For decades, patients with renal cancer have mostly undergone radical or partial nephrectomy, due to limited treatment options. However, microRNAs (miR) are becoming promising molecules for possible therapeutic purposes in several types of cancer. The miR-21 has already been associated with cell growth and apoptosis inhibition in some types of neoplasms, and in relation to renal cell carcinoma, overexpression of this has already been demonstrated, being even more relevant in the clear cell type where it has also been associated with a worse prognosis. This molecule has several target genes, among them RECK, which has the power to negatively influence MMP-9, a metalloproteinase responsible for carcinogenic processes in clear cell renal cell carcinoma. Research suggests that the CRISPR/Cas9 system can be used for mammalian gene editing and applied to basic science, biotechnology, and medicine, considering that Cas9 is compatible with different species. This system is highly efficient in introducing mutations in the precursor miR sequence, leading to loss of expression and function of interest. Therefore, we will perform miR-21 editing by CRISPR/Cas9 system in metastatic clear cell renal cell carcinoma cell line and subsequently perform the analysis of the mentioned target genes (RECK/MMP-9). Thus, considering the effective role of miRNAs on the regulation of gene expression, this pioneering study may contribute to the use of these molecules in the era of precision medicine for kidney cancer, opening a perspective of studies that may in the future favor the development of new therapeutic approaches. (AU)