Examining the Effects of the RUNX1 p.Leu43Ser Variant on FPD/AML Phenotypes Using a CRISPR/Cas9-Generated Knock-In Murine Model

Authors:
Open Access
Publisher website
Google Scholar
Abstract
Germline heterozygous variants in RUNX1 lead to Familial Platelet Disorder with Myeloid Leukemia Predisposition (FPD/AML). Cellular and/or animal models are helpful to uncovering the role of a variant in disease progression. Twenty-five mice per genotype (RUNX1WT/WT, RUNX1WT/L43S, RUNX1L43S/L43S), previously generated by CRISPR/Cas9, and nine sub-lethally irradiated mice per genotype were investigated. Peripheral blood (PB), bone marrow (BM), and spleen samples were analyzed by flow cytometry and histopathology. Deregulated genes were analyzed by RNA-seq in BM. An aberrant myeloid Mac1+Sca1+ckit− population in the PB, BM, and spleen of two homozygous and one heterozygous mouse was observed, as well as BM hypercellularity. No Mac1+Sca1+ckit− cells were detected in any RUNX1WT/WT mice. Moreover, the spleen of both homozygous mice showed destruction of the white/red pulp and the presence of apoptotic cells. The aberrant population was also detected in four irradiated mice, two heterozygous and two homozygous, in their PB, BM, and spleen. RNA-seq studies showed 698 genes significantly deregulated in the three non-irradiated Mac1+Sca1+ckit− mice vs. six healthy mice, highlighting the alteration of genes involved in apoptosis and DNA repair. These results indicate that the homozygous form of the variant p.Leu43Ser may contribute to the pathogenesis of aberrant cells.
Funding Information
  • Instituto de Salud Carlos III (ISCIII) & Feder (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • European Union (ERDF/ESF, “Investing in your future”) (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • Gerencia Regional de Salud (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • Fundación Mutua Madrileña (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • Sociedad Española de Trombosis y Hemostasia (SETH-FETH; Premio López Borrasca 2019, 2024 and Ayuda a Grupos de Trabajo en Patología Hemorrágica 2020–2024) (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • Fundación Castellano Leonesa de Hematología y Hemoterapia (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • MCIN/AEI/10.13039/501100011033 (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • ERDF “A way of making Europe” funded by the Council of Education of Junta de Castilla y León, Spain and ERDF “A way of making Europe”, Red Temática de Investigación Cooperativa en Cáncer (RTICC) (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • Centro de Investigación Biomédica en Red de Cáncer (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • “Ayuda predoctoral de la Junta de Castilla y León” by the Fondo Social Europeo (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)
  • Ayudas Postdoctorales de la Sociedad Española de Hematología y Hemoterapia (SEHH) 2022 and Ministerio de Ciencia (PI23/00624, PI23/01103, PI24/01458, PMP24/00025, GRS2551/A/22, GRS2727/A1/23, GRS2907/A1/23, AP172142019, FUCALHH 2020, PID2022-137717OB-C22, RD12/0036/0069, CIBERONC CB16/12/00233, JCYL-EDU/556/2019, JDC2023-052518-I)

This publication has 43 references indexed in Scilit: