PET-based immunomapping of intratumoral CD4+ cells to monitor acquired resistance to checkpoint inhibitors

Stefania Pezzana; Simone Blaess; Bjoern Traenkle; Anna Schaefer; Lara Ruoff; Bredi Tako; Salvador Castaneda Vega; Philipp D Kaiser; Teresa Wagner; Irene Gonzalez-Menendez; Leticia Quintanilla-Martinez; Alexander Rochwarger; Christian M Schürch; Simon Riel; Martin Schaller; Evelien A J van Genugten; Iris A E van der Hoorn; Mark A J Gorris; Megan Steinvoort; Eva Peeters; I Jolanda M de Vries; Michel M van den Heuvel; Erik H J G Aarntzen; Andreas Maurer; Ulrich Rothbauer; Bernd J Pichler; Manfred Kneilling; Dominik Sonanini

doi:10.1126/sciadv.adw1924

PET-based immunomapping of intratumoral CD4+ cells to monitor acquired resistance to checkpoint inhibitors

Stefania Pezzana, Simone Blaess, Bjoern Traenkle, Anna Schaefer, Lara Ruoff, Bredi Tako, Salvador Castaneda Vega, Philipp D Kaiser, Teresa Wagner, Irene Gonzalez-Menendez, Leticia Quintanilla-Martinez, Alexander Rochwarger, Christian M Schürch, Simon Riel, Martin Schaller, Evelien A J van Genugten, Iris A E van der Hoorn, Mark A J Gorris, Megan Steinvoort, Eva PeetersI Jolanda M de Vries, Michel M van den Heuvel, Erik H J G Aarntzen, Andreas Maurer, Ulrich Rothbauer, Bernd J Pichler, Manfred Kneilling, Dominik Sonanini^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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Abstract

CD4+ T cells are crucial in shaping response and resistance to immunotherapy. To enhance our understanding of their multifaceted functions, we developed copper-64-radiolabeled nanobodies targeting the human CD4 receptor (64Cu-CD4-Nb1) for positron emission tomography (PET). In human CD4-receptor knock-in mice, 64Cu-CD4-Nb1 specifically accumulated in different orthotopic tumors, correlating with histological CD4+ cell densities. Based on intratumoral CD4+ cell distribution patterns within the core and periphery, we distinguished responders to combined αPD-1/4-1BB antibodies early on-treatment. CD4-PET identified resistance to αPD-1 monotherapy, which was mitigated by adding regulatory T cell-depleting α4-1BB antibodies. Patients with early-stage non-small cell lung cancer who relapsed after neoadjuvant αPD-L1 therapy revealed low CD4+ T cell densities in the tumor core. In human and mouse tumor tissues, regulatory T cells correlated with CD4+ cell densities. Thus, visualizing the spatial distribution patterns of CD4+ cells by PET offers mechanistic insights into CD4-mediated therapy efficacy, with great potential for guiding combinatorial immunotherapies in patients with cancer.

Original language	English
Article number	eadw1924
Number of pages	13
Journal	Science advances
Volume	11
Issue number	26
DOIs	https://doi.org/10.1126/sciadv.adw1924
Publication status	Published - 27 Jun 2025

Keywords

Animals
CD4-Positive T-Lymphocytes/immunology
Carcinoma, Non-Small-Cell Lung/immunology
Cell Line, Tumor
Copper Radioisotopes/chemistry
Drug Resistance, Neoplasm/immunology
Humans
Immune Checkpoint Inhibitors/pharmacology
Immunotherapy
Lung Neoplasms/immunology
Mice
Positron-Emission Tomography/methods
Single-Domain Antibodies/immunology
T-Lymphocytes, Regulatory/immunology

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Pezzana, S., Blaess, S., Traenkle, B., Schaefer, A., Ruoff, L., Tako, B., Castaneda Vega, S., Kaiser, P. D., Wagner, T., Gonzalez-Menendez, I., Quintanilla-Martinez, L., Rochwarger, A., Schürch, C. M., Riel, S., Schaller, M., van Genugten, E. A. J., van der Hoorn, I. A. E., Gorris, M. A. J., Steinvoort, M., ... Sonanini, D. (2025). PET-based immunomapping of intratumoral CD4+ cells to monitor acquired resistance to checkpoint inhibitors. Science advances, 11(26), Article eadw1924. https://doi.org/10.1126/sciadv.adw1924

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title = "PET-based immunomapping of intratumoral CD4+ cells to monitor acquired resistance to checkpoint inhibitors",

abstract = "CD4+ T cells are crucial in shaping response and resistance to immunotherapy. To enhance our understanding of their multifaceted functions, we developed copper-64-radiolabeled nanobodies targeting the human CD4 receptor (64Cu-CD4-Nb1) for positron emission tomography (PET). In human CD4-receptor knock-in mice, 64Cu-CD4-Nb1 specifically accumulated in different orthotopic tumors, correlating with histological CD4+ cell densities. Based on intratumoral CD4+ cell distribution patterns within the core and periphery, we distinguished responders to combined αPD-1/4-1BB antibodies early on-treatment. CD4-PET identified resistance to αPD-1 monotherapy, which was mitigated by adding regulatory T cell-depleting α4-1BB antibodies. Patients with early-stage non-small cell lung cancer who relapsed after neoadjuvant αPD-L1 therapy revealed low CD4+ T cell densities in the tumor core. In human and mouse tumor tissues, regulatory T cells correlated with CD4+ cell densities. Thus, visualizing the spatial distribution patterns of CD4+ cells by PET offers mechanistic insights into CD4-mediated therapy efficacy, with great potential for guiding combinatorial immunotherapies in patients with cancer.",

keywords = "Animals, CD4-Positive T-Lymphocytes/immunology, Carcinoma, Non-Small-Cell Lung/immunology, Cell Line, Tumor, Copper Radioisotopes/chemistry, Drug Resistance, Neoplasm/immunology, Humans, Immune Checkpoint Inhibitors/pharmacology, Immunotherapy, Lung Neoplasms/immunology, Mice, Positron-Emission Tomography/methods, Single-Domain Antibodies/immunology, T-Lymphocytes, Regulatory/immunology",

author = "Stefania Pezzana and Simone Blaess and Bjoern Traenkle and Anna Schaefer and Lara Ruoff and Bredi Tako and \{Castaneda Vega\}, Salvador and Kaiser, \{Philipp D\} and Teresa Wagner and Irene Gonzalez-Menendez and Leticia Quintanilla-Martinez and Alexander Rochwarger and Sch{\"u}rch, \{Christian M\} and Simon Riel and Martin Schaller and \{van Genugten\}, \{Evelien A J\} and \{van der Hoorn\}, \{Iris A E\} and Gorris, \{Mark A J\} and Megan Steinvoort and Eva Peeters and \{de Vries\}, \{I Jolanda M\} and \{van den Heuvel\}, \{Michel M\} and Aarntzen, \{Erik H J G\} and Andreas Maurer and Ulrich Rothbauer and Pichler, \{Bernd J\} and Manfred Kneilling and Dominik Sonanini",

note = "Publisher Copyright: Copyright {\textcopyright} 2025 The Authors, some rights reserved;",

year = "2025",

month = jun,

day = "27",

doi = "10.1126/sciadv.adw1924",

language = "English",

volume = "11",

journal = "Science advances",

publisher = "American Association for the Advancement of Science",

number = "26",

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Pezzana, S, Blaess, S, Traenkle, B, Schaefer, A, Ruoff, L, Tako, B, Castaneda Vega, S, Kaiser, PD, Wagner, T, Gonzalez-Menendez, I, Quintanilla-Martinez, L, Rochwarger, A, Schürch, CM, Riel, S, Schaller, M, van Genugten, EAJ, van der Hoorn, IAE, Gorris, MAJ, Steinvoort, M, Peeters, E, de Vries, IJM, van den Heuvel, MM, Aarntzen, EHJG, Maurer, A, Rothbauer, U, Pichler, BJ, Kneilling, M & Sonanini, D 2025, 'PET-based immunomapping of intratumoral CD4+ cells to monitor acquired resistance to checkpoint inhibitors', Science advances, vol. 11, no. 26, eadw1924. https://doi.org/10.1126/sciadv.adw1924

TY - JOUR

T1 - PET-based immunomapping of intratumoral CD4+ cells to monitor acquired resistance to checkpoint inhibitors

AU - Pezzana, Stefania

AU - Blaess, Simone

AU - Traenkle, Bjoern

AU - Schaefer, Anna

AU - Ruoff, Lara

AU - Tako, Bredi

AU - Castaneda Vega, Salvador

AU - Kaiser, Philipp D

AU - Wagner, Teresa

AU - Gonzalez-Menendez, Irene

AU - Quintanilla-Martinez, Leticia

AU - Rochwarger, Alexander

AU - Schürch, Christian M

AU - Riel, Simon

AU - Schaller, Martin

AU - van Genugten, Evelien A J

AU - van der Hoorn, Iris A E

AU - Gorris, Mark A J

AU - Steinvoort, Megan

AU - Peeters, Eva

AU - de Vries, I Jolanda M

AU - van den Heuvel, Michel M

AU - Aarntzen, Erik H J G

AU - Maurer, Andreas

AU - Rothbauer, Ulrich

AU - Pichler, Bernd J

AU - Kneilling, Manfred

AU - Sonanini, Dominik

PY - 2025/6/27

Y1 - 2025/6/27

N2 - CD4+ T cells are crucial in shaping response and resistance to immunotherapy. To enhance our understanding of their multifaceted functions, we developed copper-64-radiolabeled nanobodies targeting the human CD4 receptor (64Cu-CD4-Nb1) for positron emission tomography (PET). In human CD4-receptor knock-in mice, 64Cu-CD4-Nb1 specifically accumulated in different orthotopic tumors, correlating with histological CD4+ cell densities. Based on intratumoral CD4+ cell distribution patterns within the core and periphery, we distinguished responders to combined αPD-1/4-1BB antibodies early on-treatment. CD4-PET identified resistance to αPD-1 monotherapy, which was mitigated by adding regulatory T cell-depleting α4-1BB antibodies. Patients with early-stage non-small cell lung cancer who relapsed after neoadjuvant αPD-L1 therapy revealed low CD4+ T cell densities in the tumor core. In human and mouse tumor tissues, regulatory T cells correlated with CD4+ cell densities. Thus, visualizing the spatial distribution patterns of CD4+ cells by PET offers mechanistic insights into CD4-mediated therapy efficacy, with great potential for guiding combinatorial immunotherapies in patients with cancer.

AB - CD4+ T cells are crucial in shaping response and resistance to immunotherapy. To enhance our understanding of their multifaceted functions, we developed copper-64-radiolabeled nanobodies targeting the human CD4 receptor (64Cu-CD4-Nb1) for positron emission tomography (PET). In human CD4-receptor knock-in mice, 64Cu-CD4-Nb1 specifically accumulated in different orthotopic tumors, correlating with histological CD4+ cell densities. Based on intratumoral CD4+ cell distribution patterns within the core and periphery, we distinguished responders to combined αPD-1/4-1BB antibodies early on-treatment. CD4-PET identified resistance to αPD-1 monotherapy, which was mitigated by adding regulatory T cell-depleting α4-1BB antibodies. Patients with early-stage non-small cell lung cancer who relapsed after neoadjuvant αPD-L1 therapy revealed low CD4+ T cell densities in the tumor core. In human and mouse tumor tissues, regulatory T cells correlated with CD4+ cell densities. Thus, visualizing the spatial distribution patterns of CD4+ cells by PET offers mechanistic insights into CD4-mediated therapy efficacy, with great potential for guiding combinatorial immunotherapies in patients with cancer.

KW - Animals

KW - CD4-Positive T-Lymphocytes/immunology

KW - Carcinoma, Non-Small-Cell Lung/immunology

KW - Cell Line, Tumor

KW - Copper Radioisotopes/chemistry

KW - Drug Resistance, Neoplasm/immunology

KW - Humans

KW - Immune Checkpoint Inhibitors/pharmacology

KW - Immunotherapy

KW - Lung Neoplasms/immunology

KW - Mice

KW - Positron-Emission Tomography/methods

KW - Single-Domain Antibodies/immunology

KW - T-Lymphocytes, Regulatory/immunology

U2 - 10.1126/sciadv.adw1924

DO - 10.1126/sciadv.adw1924

M3 - Article

C2 - 40561008

VL - 11

JO - Science advances

JF - Science advances

IS - 26

M1 - eadw1924

ER -

PET-based immunomapping of intratumoral CD4+ cells to monitor acquired resistance to checkpoint inhibitors

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