Proteomics of resistance to Notch1 inhibition in acute lymphoblastic leukemia reveals targetable kinase signatures

Giulia Franciosa; Jos G A Smits; Sonia Minuzzo; Ana Martinez-Val; Stefano Indraccolo; Jesper V Olsen

doi:10.1038/s41467-021-22787-9

Proteomics of resistance to Notch1 inhibition in acute lymphoblastic leukemia reveals targetable kinase signatures

Nat Commun. 2021 May 4;12(1):2507. doi: 10.1038/s41467-021-22787-9.

Authors

Giulia Franciosa¹, Jos G A Smits^{1

2}, Sonia Minuzzo³, Ana Martinez-Val¹, Stefano Indraccolo^{3

4}, Jesper V Olsen⁵

Affiliations

¹ Proteomics Program, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.
² Department of Molecular Developmental Biology, Radboud University, Nijmegen, Netherlands.
³ Department of Surgery, Oncology and Gastroenterology, University of Padova, Padova, Italy.
⁴ Veneto Institute of Oncology IOV - IRCCS, Padova, Italy.
⁵ Proteomics Program, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark. jesper.olsen@cpr.ku.dk.

Abstract

Notch1 is a crucial oncogenic driver in T-cell acute lymphoblastic leukemia (T-ALL), making it an attractive therapeutic target. However, the success of targeted therapy using γ-secretase inhibitors (GSIs), small molecules blocking Notch cleavage and subsequent activation, has been limited due to development of resistance, thus restricting its clinical efficacy. Here, we systematically compare GSI resistant and sensitive cell states by quantitative mass spectrometry-based phosphoproteomics, using complementary models of resistance, including T-ALL patient-derived xenografts (PDX) models. Our datasets reveal common mechanisms of GSI resistance, including a distinct kinase signature that involves protein kinase C delta. We demonstrate that the PKC inhibitor sotrastaurin enhances the anti-leukemic activity of GSI in PDX models and completely abrogates the development of acquired GSI resistance in vitro. Overall, we highlight the potential of proteomics to dissect alterations in cellular signaling and identify druggable pathways in cancer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acetophenones / pharmacology
Amyloid Precursor Protein Secretases / antagonists & inhibitors*
Amyloid Precursor Protein Secretases / metabolism
Animals
Antineoplastic Agents / pharmacology*
Antineoplastic Agents / therapeutic use
Benzopyrans / pharmacology
Cell Line, Tumor
Cell Survival / drug effects
Chromatin Immunoprecipitation
Chromatography, High Pressure Liquid
Drug Resistance, Neoplasm* / genetics
Gene Ontology
Humans
Inhibitory Concentration 50
Mice
Mice, Inbred NOD
Oligopeptides / pharmacology*
Phosphorylation
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / drug therapy*
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / metabolism*
Protein Array Analysis
Protein Biosynthesis / drug effects
Protein Kinase C / antagonists & inhibitors
Protein Kinase C / genetics
Protein Kinase C / metabolism*
Protein Kinases / metabolism
Proteomics
Receptor, Notch1 / antagonists & inhibitors*
Receptor, Notch1 / metabolism
Signal Transduction / drug effects
Tandem Mass Spectrometry
Xenograft Model Antitumor Assays

Substances

Acetophenones
Antineoplastic Agents
Benzopyrans
NOTCH1 protein, human
Oligopeptides
Receptor, Notch1
benzyloxycarbonyl-leucyl-leucyl-norleucinal
rottlerin
Protein Kinases
Protein Kinase C
Amyloid Precursor Protein Secretases