Defining the functional role of the splicing factor regulator (KIS) during leukemogenesis using a murine bone marrow transplantion model

• Grant number: 14/01458-3
• Support Opportunities: Research Grants - Young Investigators Grants
• Start date: February 01, 2015
• End date: January 31, 2020
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Leticia Fröhlich Archangelo
• Grantee: Leticia Fröhlich Archangelo
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated researchers: Alexandre Maucuer ; Jörg Kobarg ; Philipp Greif ; Sara Teresinha Olalla Saad
• Associated scholarship(s): 19/04487-8 - Consequence of G372v and T454M mutations in SF1 functionality, BP.IC; 18/07525-5 - SF1 mutations spectrum in cancer and implications on its subcellular localization and splicing function, BP.IC; 16/20071-8 - Investigation of cooperative oncogenes in IL7R-mediated acute lymphoblastic leukemia emergence, BP.MS; + associated scholarships 16/19658-4 - Functional consequence of the splicing factor SF1 inhibition in leukemic cell line, BP.MS; 16/03982-7 - Cloning SF1 wild type and mutated G372v and T454M in bicistronic retroviral vector for retrovirus production, BP.IC; 16/04372-8 - Functional characterization of KIS and its role in leukemogenesis, BP.DD; 15/22778-9 - KIS inhibitory sequences cloning in retroviral vector and stable retroviral producing lineage establishment , BP.IC - associated scholarships

Abstract

KIS (UHMK1) is a nuclear serine/threonine kinase that possesses a U2AF homology motif (UHM) and phosphorylates and regulates the activity of the splicing factors SF1 and SF3b1. Upon interaction, KIS phosphorylates SF1, which in turn enhances its binding to U2AF65 and the 3' splice site, an event known to take place during the early steps of spliceosome assembly. Somatic mutations in several components of the spliceosome machinery, including SF1 and SF3b1, have been recently implicated in leukemogenesis. The fact that KIS regulates these factors suggests that KIS might also be involved in leukemogenesis. Beside mutations, altered phosphorylation pattern of the splicing factors might also be involved in splicing deficiency and disease development. Our unpublished data showed KIS upregulation in the bone marrow of leukemia patient samples compared to controls. We hypothesized that KIS might play a role in leukemogenesis, possibly altering SF1 and SF3b1 phosphorylation. Therefore the aim of this work is to investigate the role of KIS in the leukemogenesis as well as to establish the murine bone marrow transplantation model in our institution. This mouse model is the longest and most stringent assay to test the transforming ability of an overexpressed gene at the level of the hematopoietic stem cell in the bone marrow, which mimics the leukemic process in the human the bone marrow. In order to reach our goals we intend to construct a retroviral MIY-based expression vectors (yellow fluorescent protein) of FLAG tagged KIS. MIY-KIS retrovirus will be produced to transduce primary murine bone marrow cells. Retrovirally transduced bone marrow cells, overexpressing KIS, will be transplanted into lethally irradiated syngeneic mice and used for in vitro experiments. (AU)

Scientific publications

(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)

ARFELLI, VANESSA C.; CHANG, YUN-CHIEN; BAGNOLI, JOHANNES W.; KERBS, PAUL; CIAMPONI, FELIPE E.; PAZ, LAISSA M. DA S.; PANKIVSKYI, SERHII; SALONE, JEAN DE MATHA; MAUCUER, ALEXANDRE; MASSIRER, KATLIN B.; et al. UHMK1 is a novel splicing regulatory kinase. Journal of Biological Chemistry, v. 299, n. 4, p. 22-pg., 2023-03-17. (22/08648-9, 14/01458-3, 20/02006-0)

BARBUTTI, ISABELLA; MACHADO-NETO, JOAO AGOSTINHO; ARFELLI, VANESSA CRISTINA; CAMPOS, PAULA DE MELO; TRAINA, FABIOLA; OLALLA SAAD, SARA TERESINHA; ARCHANGELO, LETICIA FROHLICH. The U2AF homology motif kinase 1 (UHMK1) is upregulated upon hematopoietic cell differentiation. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE, v. 1864, n. 3, p. 959-966, MAR 2018. (11/51959-0, 14/01458-3)