-Author name in bold denotes the presenting author
-Asterisk * with author name denotes a Non-ASH member
Clinically Relevant Abstract denotes an abstract that is clinically relevant.

PhD Trainee denotes that this is a recommended PHD Trainee Session.

Ticketed Session denotes that this is a ticketed session.

326 KRAS G12D Specific Inhibitors Relieve Erythroid Differentiation Block and Modulate Inflammatory Pathways at the Single Cell Level in Myeloid Malignancies

Program: Oral and Poster Abstracts
Type: Oral
Session: 618. Acute Myeloid Leukemias: Biomarkers and Molecular Markers in Diagnosis and Prognosis: Functional Genomics in Prognosis and Novel Therapies
Hematology Disease Topics & Pathways:
Acute Myeloid Malignancies, Research, MDS, AML, Translational Research, CHIP, Chronic Myeloid Malignancies, Hematopoiesis, Immune mechanism, Diseases, Immunology, Biological Processes, Myeloid Malignancies
Saturday, December 7, 2024: 4:15 PM

Leah Kravets, MS1, Mirca S. Saurty-Seerunghen, PhD2*, Divij Verma, PhD1, Ariel Fromowitz, MD3, Srabani Sahu, MS1*, Bradley Rockwell, MD4, Olivia Sakaguchi2*, Swathi Narayanagari, MS5*, Shanisha Gordon Mitchell1*, Milagros Carbajal1*, Srinivas Aluri, PhD1, Jingli Wang, PhD5*, Lindsay Lafave, PhD5*, Marina Konopleva6, Anna S. Nam, MD2 and Amit Verma, MD1

1Blood Cancer Institute, Department of Oncology, Albert Einstein College of Medicine, Bronx, NY
2Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York, NY
3Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX
4Blood Cancer Institute, Department of Oncology, Albert Einstein College of Medicine, New York, NY
5Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY
6Montefiore Einstein Comprehensive Cancer Center, Albert Einstein College of Medicine, Bronx, NY

Introduction:

Myelodysplastic Syndrome (MDS) and Acute Myeloid Leukemia (AML) are characterized by a hematopoietic differentiation block that leads to anemia, a major source of morbidity and mortality. These myeloid malignancies and clonal hematopoiesis (CH) are associated with a myeloid bias and heightened inflammation, but the mechanisms of these phenomenon are not well elucidated. KRAS G12D (KRASG12D) mutation is seen in ~5% of MDS/AML and associated with a poor prognosis, currently with no approved precision medicine strategies. Novel KRASG12D inhibitors are being developed for solid tumors, but no data exists testing their efficacy in myeloid malignancies. Thus, we examined the impact of KRASG12D mutation and KRAS inhibition on human hematopoietic differentiation.

Results:

To first define the impact of KRASG12D on human hematopoiesis, we performed single-cell RNA-seq coupled with mutational calling via Genotyping of Transcriptomics (GoT) on primary KRASG12D samples. GoT of a CH sample with isolated KRASG12D revealed that the KRASG12D mutated (MT) cells were overrepresented in the monocytic compartment. MT cells also displayed an inflammatory signature, including elevated IL-8 expression.

GoT analysis of KRASG12D AML samples (N=3) revealed that the MT cells were enriched in a distinct AML subpopulation, defined by a stem/progenitor-like expression profile, with upregulated RUNX1, ETV6, JARID2, and ZEB2. Additionally, CD83, an immune checkpoint molecule, was elevated in this AML subgroup. Differential gene expression analysis between CD83+ MT vs. CD83+ WT cells revealed overexpression of IL-6, FCN1, CXCL3 in MT cells, suggesting an inflamed phenotype. Consistent with CD83 upregulation in the stem/progenitor-like subgroup, sorted CD83+ cells from KRASG12D primary MDS samples create more colonies in vitro when compared to sorted CD83- cells.

To test the impact of KRAS-targeted therapy on isolated KRASG12D clones (without confounding co-occurring mutations), we treated the KRASG12D CH sample with a clinically valid KRASG12D inhibitor MRTX1133 and a pan-KRAS inhibitor BI-2865. Treatment with both inhibitors decreased total monocytic population while increasing erythrocytes at the single cell level. We validated this finding by treating primary KRASG12D mutant CH, MDS and AML samples (N=3) with both KRAS-inhibitors in clonogenic assays. Inhibition of KRASG12D led to increased erythroid colonies, associated with increased Glycophorin A and CD71 expression by flow cytometry. This demonstrates that KRAS-inhibition may rescue erythroid differentiation block.

As we observed that KRASG12D induced inflammatory signaling, we hypothesized that KRAS inhibition may rescue erythroid differentiation by suppressing inflammation. Consistently, the KRASG12D-induced inflammatory signature, including IL-8, was downregulated upon treatment with MRTX1133 and BI-2865. We therefore tested whether IL-8 antibody treatment may rescue erythroid differentiation. Indeed, IL-8 antibody in vitro treatment led to increased Glycophorin A and CD71 expression, suggesting that KRAS inhibition may rescue KRASG12D -mediated anemia via downregulation of IL-8-mediated inflammation.

Conclusion:

These data demonstrate that KRASG12D induces a monocytic differentiation bias associated with inflammation in CH. In AML, KRAS mutant cells are enriched in a novel stem progenitor-like inflamed population, characterized by CD83 positivity. We demonstrate the efficacy of novel, clinically relevant KRASG12D inhibitors in relieving IL8-mediated inflammation and erythroid differentiation block in KRASG12D neoplasms. These data support investigations of KRASG12D inhibitors in myeloid malignancy clinical trials. Ongoing studies test the efficacy of in vivo targeting of KRASG12D clones in murine and xenograft models.

Disclosures: Konopleva: Sanofi Aventis: Consultancy; Curis: Consultancy; Servier: Speakers Bureau; Intellisphere: Speakers Bureau; Novartis: Membership on an entity's Board of Directors or advisory committees; Auxenion GmbH: Membership on an entity's Board of Directors or advisory committees; Syndax: Membership on an entity's Board of Directors or advisory committees; Adaptive: Consultancy; MEI Pharma: Consultancy, Membership on an entity's Board of Directors or advisory committees; Legend Biotech: Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy, Other: clinical trials; AbbVie: Consultancy, Membership on an entity's Board of Directors or advisory committees, Other: clinical trials, Research Funding; Dark Blue Therapeutics: Membership on an entity's Board of Directors or advisory committees; Menarini Group: Consultancy, Membership on an entity's Board of Directors or advisory committees; Klondike Biopharma: Research Funding; Vincerx: Consultancy. Verma: Stelexis: Current equity holder in private company, Membership on an entity's Board of Directors or advisory committees; Clinstreet: Current equity holder in private company; Bioconvergent health: Current equity holder in private company; Prelude: Research Funding; Bristol Myers Squib: Research Funding; Curis: Membership on an entity's Board of Directors or advisory committees, Research Funding; Halia: Research Funding; Calico: Membership on an entity's Board of Directors or advisory committees.

*signifies non-member of ASH