Dynamic genetic and nongenetic RAS pathway activation drives resistance to FLT3 and BCL2 inhibitor therapy

V Vanessa E Kennedy (Stanford University, Stanford, California, United States) C Cheryl A. C. Peretz (University of California San Francisco, San Francisco, California, United States) A Anushka Walia (Brigham and Women's Hospital, Boston, Massachusetts, United States) B Brenda Chyla (Abbvie, North Chicago, Illinois, United States) Y Yan Sun J Jason E. Hill (Astellas Pharma, Northbrook, Illinois, United States) E Elaine Tran (University of California San Francisco, San Francisco, California, United States) A Andrew D. Koh (University of California San Francisco, San Francisco, California, United States) T Timothy T Ferng (University of California (San Francisco), San Francisco, California, United States) S Samantha Pintar (University of California San Francisco, San Francisco, California, United States) M Matthew Jones B Bogdan Popescu (University of California San Francisco, San Francisco, California, United States) I Isabelle Lomeli (University of California San Francisco, San Francisco, California, United States) F Farid Chehab (University of California, San Francisco) N Natalia Murad (University of California San Francisco, San Francisco, California, United States) A August John (University of California San Francisco, San Francisco, California, United States) R Ritu Parna Roy (University of California San Francisco, San Francisco, California, United States) A Adam B. Olshen (UCSF, San Francisco, California, United States) C Christine A. Berryhill (Indiana University, Indianapolis, Indiana, United States) C Christopher Davis (Indiana University School of Medicine, Department of Pediatrics, Riley Hospital for Children at IU Health) S Steven Patrick Angus (Indiana University School of Medicine, Indianapolis, Indiana, United States) J Jose M. Rivera (Benioff Children's Hospital, University of California, San Francisco, San Francisco, California, United States) A Alicia Meshulam (University of California, San Francisco, San Francisco, California, United States) E Elliot Stieglitz S Sunil Kumar Joshi E Elie Traer (Oregon Health & Science University, Portland, Oregon, United States) M Monique Dail (Illuminate Biosciences, Moss Beach, California, United States) H Habib Hamidi J Jessica K Altman (Northwestern University, Chicago, Illinois, United States) N Naval G Daver (University of Texas, MD Anderson Cancer Center, Houston, Texas, United States) M Mark J. Levis (Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine) J James McCloskey (John Theurer Cancer Center at Hackensack University Medical Center, Hackensack, New Jersey, United States) A Alexander E. Perl (Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania, United States) C Catherine C. Smith (University of California (San Francisco), San Francisco, California, United States)

Abstract

Bulk sequencing of relapsed tumors reveals mutations associated with resistance to cancer therapy but is insufficient to fully assess all causes of relapse. Due to inherent tumor heterogeneity, on-treatment tumor evolution may select for genetically distinct clones or shifts in malignant transcriptional states not resolvable by bulk sequencing. We performed multiomic single cell (SC) DNA/protein and RNA/protein profiling of a clinical trial cohort of acute myeloid leukemia (AML) patients treated on the Phase 1b clinical trial of the BCL2 inhibitor venetoclax and the FLT3 inhibitor gilteritinib (Ven/Gilt) to characterize immunophenotypic, transcriptional, and genetic clonal evolution driving resistance. We found that while Ven/Gilt effectively eliminated FLT3 mutant clones, resistance was associated with RAS activation via multiple mechanisms including selection for RAS mutant clones, non-mutational upregulation of RAS transcriptional programs and a shift to RAS-associated monocytic AML differentiation. In an in vitro model of monocytic differentiation associated with non-mutational RAS transcriptional activation, we demonstrated that RAS pathway inhibition re-sensitized to Ven/Gilt. These data illustrate that convergent resistance pathways in patients can be activated via diverse genetic and non-genetic mechanisms. These results underscore that RAS signaling is central to FLT3 and BCL2 inhibitor resistance, is tightly coupled to AML monocytic differentiation and highlight RAS pathway inhibition as a viable clinical strategy to combat resistance. CT# NCT03625505

Article Details

Journal Blood
Volume / Issue Vol. 1, Issue 1
Published June 01, 2026
ISSN 0006-4971
Publisher Elsevier BV

Journal Info

Blood

Elsevier BV

ISSN: 0006-4971 Health Sciences

Authors (34)

V

Vanessa E Kennedy

Stanford University, Stanford, California, United States

C

Cheryl A. C. Peretz

University of California San Francisco, San Francisco, California, United States

A

Anushka Walia

Brigham and Women's Hospital, Boston, Massachusetts, United States

B

Brenda Chyla

Abbvie, North Chicago, Illinois, United States

Y

Yan Sun

J

Jason E. Hill

Astellas Pharma, Northbrook, Illinois, United States

E

Elaine Tran

University of California San Francisco, San Francisco, California, United States

A

Andrew D. Koh

University of California San Francisco, San Francisco, California, United States

T

Timothy T Ferng

University of California (San Francisco), San Francisco, California, United States

S

Samantha Pintar

University of California San Francisco, San Francisco, California, United States

M

Matthew Jones

B

Bogdan Popescu

University of California San Francisco, San Francisco, California, United States

I

Isabelle Lomeli

University of California San Francisco, San Francisco, California, United States

F

Farid Chehab

University of California, San Francisco

N

Natalia Murad

University of California San Francisco, San Francisco, California, United States

A

August John

University of California San Francisco, San Francisco, California, United States

R

Ritu Parna Roy

University of California San Francisco, San Francisco, California, United States

A

Adam B. Olshen

UCSF, San Francisco, California, United States

C

Christine A. Berryhill

Indiana University, Indianapolis, Indiana, United States

C

Christopher Davis

Indiana University School of Medicine, Department of Pediatrics, Riley Hospital for Children at IU Health

S

Steven Patrick Angus

Indiana University School of Medicine, Indianapolis, Indiana, United States

J

Jose M. Rivera

Benioff Children's Hospital, University of California, San Francisco, San Francisco, California, United States

A

Alicia Meshulam

University of California, San Francisco, San Francisco, California, United States

E

Elliot Stieglitz

S

Sunil Kumar Joshi

E

Elie Traer

Oregon Health & Science University, Portland, Oregon, United States

M

Monique Dail

Illuminate Biosciences, Moss Beach, California, United States

H

Habib Hamidi

J

Jessica K Altman

Northwestern University, Chicago, Illinois, United States

N

Naval G Daver

University of Texas, MD Anderson Cancer Center, Houston, Texas, United States

M

Mark J. Levis

Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine

J

James McCloskey

John Theurer Cancer Center at Hackensack University Medical Center, Hackensack, New Jersey, United States

A

Alexander E. Perl

Abramson Cancer Center, University of Pennsylvania, Philadelphia, Pennsylvania, United States

C

Catherine C. Smith

University of California (San Francisco), San Francisco, California, United States