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    Poster Networking

    Coagulation and Natural Anticoagulants

    Poster Networking Session

    PB0525 - Identification of an aberrant cell population in a knock-in murine model with RUNX1 variant generated by CRISPR/Cas9

    Monday, June 26, 2023
    18:30 – 19:30 ET
    Room: Exhibition

      Presenting Author(s)

    • AM

      Ana Marin-Quilez, PhD

      Postdoctoral researcher
      Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, University of Murcia, IMIB, CIBERER-U765, Murcia, Spain
      Murcia, Murcia, Spain

      • Co-Author(s)

    • IG

      Ignacio García-Tuñón

      Professor
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain
      Salamanca, Castilla y Leon, Spain

    • JO

      José Luis Ordoñez

      Proffesor
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain; Laboratory of Pharmacology, Department of Physiology and Pharmacology, Faculty of Pharmacy, University of Salamanca, Salamanca, Spain
      Salamanca, Castilla y Leon, Spain

    • Md

      Mónica del Rey

      Postdoctoral researcher
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain
      Salamanca, Castilla y Leon, Spain

    • DS

      David Sanz

      Postdoctoral researcher
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain
      Salamanca, Castilla y Leon, Spain

    • LD

      Lorena Díaz-Ajenjo

      Predoctoral researcher
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain
      Salamanca, Castilla y Leon, Spain

    • CG

      Carmen Guerrero

      Principal Investigator
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain
      Salamanca, Castilla y Leon, Spain

    • JP

      Jesús Pérez-Losada

      Principal Investigator
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain
      Salamanca, Castilla y Leon, Spain

    • JG

      Jose Ramon Gonzalez Porras

      Hematologo
      Hospital Clínico Universitario Salamanca
      salamanca, Castilla y Leon, Spain

    • JH

      Jesús María Hernández-Rivas

      Principal Investigator
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain; Hospital Clínico Universitario Salamanca
      Salamanca, Castilla y Leon, Spain

    • RB

      Rocío Benito

      Postdoctoral researcher
      Cancer Research Center - CSIC, University of Salamanca, Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain
      Salamanca, Castilla y Leon, Spain

    • José RIVERA Rivera, PhD

      PhD
      Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, Murcia, Spain
      Murcia, Murcia, Spain

    • Jose Mª Bastida, PhD, MD

      Hematology Department
      Complejo Asistencial Universitario de Salamanca-IBSAL-USAL (Salamanca)
      Salamanca, Castilla y Leon, Spain

    Background: Germline heterozygous variants in RUNX1 led to the Familial Platelet Disorder with Myeloid Leukemia Predisposition (FPD/AML). Although an acquired second oncogenic hit is needed to develop a myeloid neoplasia, the mechanisms are not well understood. Cellular and/or animal models provide a useful tool to uncover the potential role of RUNX1 in the disease progression.

    Aims: Characterization of leukemogenesis in a CRISPR/Cas9-generated knock-in murine model carrying the germline p.Leu43Ser variant in RUNX1 (mimicking human p.Leu56Ser), and identification of second events and transcriptionally deregulated genes.

    Methods: 75 mice were studied: 25 of each genotype (RUNX1WT/WT, RUNX1WT/L43S, RUNX1L43S/L43S). Peripheral blood (PB), bone marrow (BM), and spleen were analyzed by flow cytometry and histopathology. To detect somatic events, all exons of Runx1 were sequenced, and gene expression profile of 3 mice/genotype was analyzed by RNA-seq in BM.

    Results: We detected the presence of aberrant myeloid Mac1+Sca1+ckit- cells in PB of two homozygous mice (Hom#1 at 15 months, and Hom#2 at 20 months), and in only one heterozygous mouse (Het#1 at 21 months of age). These aberrant cells were detected in both BM and spleen (Table 1). No Mac1+Sca1+ckit- cells were observed in any RUNX1WT/WT mice. BM hypercellularity was detected in the 3 Mac1+Sca1+ckit- mice. Moreover, the spleen of Hom#1 and Hom#2 mice showed a destruction of the white/red pulp and the presence of apoptotic cells (Figure 1). DNA-seq analyses showed no additional somatic variants in Runx1. However, RNA-seq studies showed 698 genes significantly downregulated in the 3 Mac1+Sca1+ckit- mice vs. 6 healthy mice, highlighting the alteration of genes involved in apoptosis and DNA repair.

    Conclusion(s): We have identified aberrant cells in two mice with the RUNX1 p.Leu43Ser variant in homozygosis and in one heterozygous mouse. These results suggest that the variant p.Leu43Ser is associated with an aberrant cell phenotype, being milder in the heterozygous state.

    Funding: PI20/00926, 21920/PI/22.