The role of NEK1 loss-of-function ALS mutations for oxidative stress, DNA damage and cell death

• Grant number: 21/14661-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: April 15, 2022
• End date: December 31, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Jörg Kobarg
• Grantee: Luidy Kazuo Issayama
• Supervisor: David M. Wilson III
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Institution abroad: Hasselt University, Diepenbeek, Belgium
• Associated to the scholarship: 18/13775-4 - The human protein kinase NEK1: the characterization of its physiological activation by DNA damage and its use as a biological marker and potential therapeutic target in Thyroid Cancer, BP.DD

Abstract

NEK1 has pleiotropic functions and mutations of its gene have been reported in cancer and ciliopathies, such as polycystic kidney disease (PKD), besides some genetic diseases such as Mohr syndrome and short-rib thoracic dysplasia (SRTD). Recent studies demonstrate that loss-of-function mutations of NEK1 are related to familial ALS, however, how those mutations contribute to development of the disease is not fully understood. Our preliminary results suggest that NEK1 knockout HEK293T cells increase reactive oxygen species (ROS), which contributes to DNA damage accumulation and leads to cell death. Additionally, when treated with etoposide, a chemotherapeutic agent that inhibits topoisomerase II and induces double strand breaks (DSBs), NEK1 knockout cells keeps expressing ³-H2AX, a molecular marker for DSBs, after 240 minutes of recovery (after treatment). In this project, we aim to study and validate interactions of key components for DNA damage repair mechanisms and mitochondrial activity (such as MRE11, VHL and VDAC1). Additionally, by using HEK293T NEK1 knockout cells and transfecting NEK1 mutated constructions we aim to understand the direct effect of those mutations on interactors or cell physiology. We believe that Dr. David M. Wilson III and his network (Hässelt University) expertise in DNA damage, oxidative stress and neurodegenerative diseases will provide better understanding for NEK1 physiological role. (AU)