From functional studies to searching for new inhibitors for cancer: exploring kinases that regulate the cell cycle of the human NEK family

• Grant number: 17/03489-1
• Support Opportunities: Research Projects - Thematic Grants
• Start date: December 01, 2017
• End date: April 30, 2023
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Jörg Kobarg
• Grantee: Jörg Kobarg
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated researchers: Carlos Frederico Martins Menck ; Daniel Fábio Kawano ; Daniel Martins-de-Souza ; Guido Lenz ; Hernandes Faustino de Carvalho ; Jonathan Mark Elkins ; Leonardo dos Reis Silveira ; Marcelo Bispo de Jesus ; Vadim Viviani
• Associated scholarship(s): 21/09439-1 - Investigation of the regulatory role of Nek4 in the alternative splicing in non-small cell lung cancer, BP.MS; 21/12888-2 - From functional studies to a search for new anti-cancer inhibitors: exploring the cell cycle regulating kinases of the human NEK family, BP.TT; 20/16265-7 - Identification and functional studies of NEK1 and NEK2 interaction partners and substrates on the human centrosome and their relevance in ciliopathies, BP.PD; + associated scholarships 20/07006-8 - Analysis of the role of Nek10 in mitochondria, BP.PD; 19/25188-9 - Evaluation of the involvement of the human kinases NEK4 and Nek5 in the intrinsic apoptosis pathway After DNA damage, BP.IC; 18/25614-5 - Analysis of the role of the Nek family members in adjusting the response to DNA damage in mitochondria, BP.DD; 18/08391-2 - Functional and molecular studies of the human protein kinase Nek6 as a target candidate for drug design in Prostate Cancer, BP.DR; 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; 18/05350-3 - Study of the role of the interaction between mitofusin and Nek4 in the signaling between mitochondria and nucleus after cellular stress, BP.PD; 15/06458-4 - Functional characterization of regulatory proteins involved in DNA repair, BP.PD - associated scholarships

Abstract

Members of the family of NEK kinases (NIMA related kinases) were identified as important regulators of cell cycle checkpoints, especially at the G2 to M phase transition. Although they are among all kinases one of the least studied families, recent studies showed they can have crucial roles in mitosis, centrosome disjunction, and in the signaling of the DNA damage response. These characteristics along with the fact that several NEKs were found to be over-expressed in cancer, or were shown to present elevated mutation rates, suggest that they are interesting candidates both in the diagnostic as well as in the therapy of cancer. However, for most NEKs the physiological substrates have not been identified and nor are the functional consequences of the found point mutations known that occur in their genes in cancer cells. Therefore, the central aim of this project is to elucidate the functional roles of the NEKs in normal and tumoral cells. We will use the "Shokat" approach to generate kinase that are sensitive to ATP analogs to identify physiological substrates of the NEKs. Furthermore, we will generate mutations observed in Nek genes in cancer tissues to analyze functional consequences of these altered kinases in vitro and in vivo. Furthermore, we will characterize the protein expression of NEKs in normal and tumor tissues and 'high throughput screening' of inhibitors, based on coupled kinase/luciferase ATP consumption bioassay, previously developed in our group. In summary, our project envisions to study the physiological and pathophysiological roles of NEK 1, 3, 4, 5, 6, 7, 8 and 10. The studies will contribute to explore the potential of these cell cycle regulatory kinases as novel targets in the therapy of cancer and envision the discovery of novel anti-cancer inhibitors. (AU)

Scientific publications

(13)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

BASEI, FERNANDA L.; MOURA, LIVIA A. R.; KOBARG, JORG. Using the E1A Minigene Tool to Study mRNA Splicing Changes. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, n. 170, APR 2021. (17/03489-1, 18/05350-3)

PERES DE OLIVEIRA, ANDRESSA; BASEI, FERNANDA LUISA; SLEPICKA, PRISCILA FERREIRA; DE CASTRO FEREZIN, CAMILA; MELO-HANCHUK, TALITA D.; DE SOUZA, EDMARCIA ELISA; LIMA, TANES I.; DOS SANTOS, VALQUIRIA TIAGO; MENDES, DAVI; SILVEIRA, LEONARDO REIS; et al. NEK10 interactome and depletion reveal new roles in mitochondria. PROTEOME SCIENCE, v. 18, n. 1, APR 28 2020. (14/15982-6, 18/05350-3, 17/03489-1)

DE OLIVEIRA, ANDRESSA PERES; ISSAYAMA, LUIDY KAZUO; BETIM PAVAN, ISADORA CAROLINA; SILVA, FERNANDO RIBACK; MELO-HANCHUK, TALITA DINIZ; SIMABUCO, FERNANDO MOREIRA; KOBARG, JORG. Checking NEKs: Overcoming a Bottleneck in Human Diseases. Molecules, v. 25, n. 8, APR 2 2020. (14/15982-6, 18/14818-9, 17/03489-1)

BASEI, FERNANDA LUISA; FEREZIN, CAMILA DE CASTRO; RODRIGUES DE OLIVEIRA, ANA LUISA; MUNOZ, JUAN PABLO; ZORZANO, ANTONIO; KOBARG, JORG. Nek4 regulates mitochondrial respiration and morphology. FEBS Journal, v. 289, n. 11, p. 18-pg., 2022-01-24. (18/05350-3, 17/03489-1)

MELO-HANCHUK, TALITA DINIZ; SLEPICKA, PRISCILA FERREIRA; PELEGRINI, ALESSANDRA LUIZA; MARTINS MENCK, CARLOS FREDERICO; KOBARG, JORG. NEK5 interacts with topoisomerase II beta and is involved in the DNA damage response induced by etoposide. Journal of Cellular Biochemistry, v. 120, n. 10, p. 16853-16866, OCT 2019. (14/15982-6, 17/03489-1, 10/51730-0, 15/06458-4, 10/16831-0, 10/15262-2)

FEREZIN, CAMILA DE CASTRO; BASEI, FERNANDA LUISA; MELO-HANCHUK, TALITA D.; DE OLIVEIRA, ANA LUISA; PERES DE OLIVEIRA, ANDRESSA; MORI, MATEUS P.; DE SOUZA-PINTO, NADJA C.; KOBARG, JORG. NEK5 interacts with LonP1 and its kinase activity is essential for the regulation of mitochondrial functions and mtDNA maintenance. FEBS OPEN BIO, v. 11, n. 3, FEB 2021. (16/10530-5, 17/03489-1, 18/05350-3, 14/50938-8)

MARTINS, MARIANA BONJIORNO; PEREZ, ARINA MARINA; BOHR, VILHELM A.; WILSON, III, DAVID M.; KOBARG, JORG. NEK1 deficiency affects mitochondrial functions and the transcriptome of key DNA repair pathways. MUTAGENESIS, v. 36, n. 3, p. 223-236, MAY 2021. (16/02040-8, 14/15982-6, 17/21067-7, 17/03489-1)

MELO-HANCHUK, TALITA DINIZ; MARTINS, MARIANA BONJIORNO; CUNHA, LUCAS LEITE; SOARES, FERNANDO AUGUSTO; WARD, LAURA STERIAN; VASSALLO, JOSE; KOBARG, JORG. Expression of the NEK family in normal and cancer tissue: an immunohistochemical study. BMC CANCER, v. 20, n. 1, JAN 6 2020. (15/06458-4, 17/03489-1, 16/02040-8)

ALVES BARBOSA, EVERTON DE ALMEIDA; SERAPHIM, THIAGO VARGAS; GANDIN, CESAR AUGUSTO; TEIXEIRA, LEILANE FERREIRA; GONCALVES DA SILVA, RONNI ANDERSON; RIGHETTO, GERMANNA L.; GONCALVES, KALIANDRA DE ALMEIDA; VASCONCELLOS, RAPHAEL DE SOUZA; ALMEIDA, MARCIA ROGERIA; SILVA JUNIOR, ABELARDO; et al. Insights into the full-length SRPK2 structure and its hydrodynamic behavior. International Journal of Biological Macromolecules, v. 137, p. 205-214, SEP 15 2019. (14/07206-6, 13/50724-5, 11/23110-0, 12/00195-3, 17/03489-1, 12/50161-8, 17/07335-9)

BETIM PAVAN, ISADORA CAROLINA; DE OLIVEIRA, ANDRESSA PERES; FIRMINO DIAS, PEDRO RAFAEL; BASEI, FERNANDA LUISA; ISSAYAMA, LUIDY KAZUO; FEREZIN, CAMILA DE CASTRO; SILVA, FERNANDO RIBACK; RODRIGUES DE OLIVEIRA, ANA LUISA; ALVES DOS REIS MOURA, LIVIA; MARTINS, MARIANA BONJIORNO; et al. On Broken Ne(c)ks and Broken DNA: The Role of Human NEKs in the DNA Damage Response. CELLS, v. 10, n. 3, MAR 2021. (16/02040-8, 17/21067-7, 17/03489-1)

FEREZIN, CAMILA DE CASTRO; SIAN, TERRY C. C. LIM KAM; WU, YUNJIAN; MA, XIUQUAN; CHUEEH, ANDERLY C.; HUANG, CHENG; SCHITTENHELM, RALF B.; KOBARG, JORG; DALY, ROGER J.. Identification of biological pathways and processes regulated by NEK5 in breast epithelial cells via an integrated proteomic approach. CELL COMMUNICATION AND SIGNALING, v. 20, n. 1, p. 15-pg., 2022-12-22. (16/10530-5, 19/11435-4, 17/03489-1)

BASEI, FERNANDA L.; MOURA, LIVIA A. R.; KOBARG, JORG. Using the E1A Minigene Tool to Study mRNA Splicing Changes. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, v. N/A, n. 170, p. 13-pg., 2021-04-01. (17/03489-1, 18/05350-3)

CASTELUCCI, B. G.; PEREIRA, A. H. M.; FIORAMONTE, M.; CARAZZOLLE, M. F.; DE OLIVEIRA, P. S. L.; FRANCHINI, K. G.; KOBARG, J.; MARTINS-DE-SOUZA, D.; JOAZEIRO, P. P.; CONSONNI, S. R.. Evidence of macrophage modulation in the mouse pubic symphysis remodeling during the end of first pregnancy and postpartum. SCIENTIFIC REPORTS, v. 10, n. 1, JUL 24 2020. (15/23616-2, 17/03489-1)