Single-cell characterization of anti–LAG-3 and anti–PD-1 combination treatment in patients with melanoma

Clinical MedicineImmunologyOncology Open Access | 10.1172/JCI164809

Jani Huuhtanen,1,2,3,4 Henna Kasanen,1,2,4 Katriina Peltola,4,5 Tapio Lönnberg,6 Virpi Glumoff,7 Oscar Brück,1,2,4 Olli Dufva,1,2,4 Karita Peltonen,1,2,4 Johanna Vikkula,3 Emmi Jokinen,1,2,3,4 Mette Ilander,1,2 Moon Hee Lee,1,2,4 Siru Mäkelä,5 Marta Nyakas,8 Bin Li,9 Micaela Hernberg,5 Petri Bono,5 Harri Lähdesmäki,3 Anna Kreutzman,1,2 and Satu Mustjoki1,2,4

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Huuhtanen, J. in: JCI | PubMed | Google Scholar |

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Kasanen, H. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Peltola, K. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Lönnberg, T. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Glumoff, V. in: JCI | PubMed | Google Scholar |

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Brück, O. in: JCI | PubMed | Google Scholar |

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Dufva, O. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Peltonen, K. in: JCI | PubMed | Google Scholar |

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Vikkula, J. in: JCI | PubMed | Google Scholar |

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Jokinen, E. in: JCI | PubMed | Google Scholar |

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Ilander, M. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Lee, M. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Mäkelä, S. in: JCI | PubMed | Google Scholar |

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Nyakas, M. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Li, B. in: JCI | PubMed | Google Scholar |

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Hernberg, M. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Bono, P. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Lähdesmäki, H. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Kreutzman, A. in: JCI | PubMed | Google Scholar

1Translational Immunology Research Program and Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki, Finland.

2Hematology Research Unit Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

3Department of Computer Science, Aalto University, Espoo, Finland.

4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland.

5Department of Oncology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.

6Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.

7Research Unit of Biomedicine, Medical Microbiology and Immunology, University of Oulu, Oulu, Finland.

8Oslo University Hospital-Radiumhospitalet, Oslo, Norway.

9Bristol Myers Squibb (BMS) Research and Development, Princeton, New Jersey, USA.

Address correspondence to: Satu Mustjoki, Hematology Research Unit Helsinki, University of Helsinki and, Helsinki University Hospital Comprehensive Cancer Center. Haartmaninkatu 8, FIN-00290 Helsinki, Finland. Phone: 358.9.471.71898; Email: satu.mustjoki@helsinki.fi.

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Find articles by Mustjoki, S. in: JCI | PubMed | Google Scholar |

Authorship note: JH and HK are co–first authors. AK and S Mustjoki are co–senior authors.

Published January 31, 2023 - More info

Published in Volume 133, Issue 6 on March 15, 2023
J Clin Invest. 2023;133(6):e164809. https://doi.org/10.1172/JCI164809.
© 2023 Huuhtanen et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Published January 31, 2023 - Version history
Received: August 25, 2022; Accepted: January 25, 2023 View PDF Abstract

Background. Relatlimab plus nivolumab (anti–lymphocyte-activation gene 3 plus anti–programmed death 1 [anti–LAG-3+anti–PD-1]) has been approved by the FDA as a first-line therapy for stage III/IV melanoma, but its detailed effect on the immune system is unknown.

Methods. We evaluated blood samples from 40 immunotherapy-naive or prior immunotherapy–refractory patients with metastatic melanoma treated with anti–LAG-3+anti–PD-1 in a phase I trial using single-cell RNA and T cell receptor sequencing (scRNA+TCRαβ-Seq) combined with other multiomics profiling.

Results. The highest LAG3 expression was noted in NK cells, Tregs, and CD8+ T cells, and these cell populations underwent the most significant changes during the treatment. Adaptive NK cells were enriched in responders and underwent profound transcriptomic changes during the therapy, resulting in an active phenotype. LAG3+ Tregs expanded, but based on the transcriptome profile, became metabolically silent during the treatment. Last, higher baseline TCR clonality was observed in responding patients, and their expanding CD8+ T cell clones gained a more cytotoxic and NK-like phenotype.

Conclusion. Anti–LAG-3+anti–PD-1 therapy has profound effects on NK cells and Tregs in addition to CD8+ T cells.

Trial registration. ClinicalTrials.gov (NCT01968109)

Funding. Cancer Foundation Finland, Sigrid Juselius Foundation, Signe and Ane Gyllenberg Foundation, Relander Foundation, State funding for university-level health research in Finland, a Helsinki Institute of Life Sciences Fellow grant, Academy of Finland (grant numbers 314442, 311081, 335432, and 335436), and an investigator-initiated research grant from BMS.

Graphical Abstractgraphical abstract Introduction

Even though immune checkpoint inhibitor therapies have revolutionized the treatment of metastatic melanoma, a majority of patients fail to achieve sustainable responses. As currently available immune checkpoint inhibitor therapies (anti–cytotoxic T lymphocyte–associated protein 4 [anti–CTLA-4], anti–programmed death 1 [anti–PD-1], and anti–programmed death ligand 1 [anti–PD-L1] therapies) primarily target effector CD8+ T cells, novel combination treatments that could also invigorate other immune cell types could increase the response rates in patients. Lymphocyte-activation gene 3 (LAG-3) is an inhibitory receptor expressed widely on different activated and exhausted immune cell subtypes (17), rendering it one of the most interesting novel immune checkpoint targets. Coinhibition of anti–LAG-3+anti–PD-1 is more attractive than blocking either LAG-3 or PD-1 alone (8), with encouraging efficacy even in patients with anti–PD-1/anti–PD-L1–refractory melanoma (9, 10). Relatlimab plus nivolumab (anti–LAG-3+anti–PD-1) combination therapy has shown a progression-free survival benefit over anti–PD-1 monotherapy as a first-line treatment for patients with metastatic melanoma (11) and has now been approved by the FDA. Although it is known that LAG-3 attenuates T cell activation, viability, and proliferation by binding to MHC class II molecules, knowledge of its effects on other immune cells is lagging (8, 1215).

In this study, we used single-cell RNA and T cell receptor (TCR) sequencing (scRNA+TCRαβ-Seq), flow cytometry, TCRβ-Seq, and serum protein profiling together with ex vivo functional validations to analyze immune cell responses to anti–LAG-3+anti–PD-1 treatment (relatlimab+nivolumab, phase I, ClinicalTrials.gov NCT01968109) in pretreatment blood samples and blood samples taken 1 and 3 months after therapy from 40 patients with metastatic melanoma (Figure 1). The patients were either immunotherapy naive (IO naive) or prior immunotherapy refractory (IO refractory) (patient details are provided in Table 1 and Supplemental Table 1; supplemental material available online with this article; https://doi.org/10.1172/JCI164809DS1), and during the therapy more changes were observed in the immune cell repertoire of IO-naive patients. Anti–LAG-3+anti–PD-1 shifted LAG-3+CD8+ antigen-experienced T cells from an exhausted to a more cytotoxic phenotype. However, we observed the greatest effect in CMV seropositivity–associated cell populations, such as in adaptive NK cells, resulting in an activated phenotype. This was mainly observed in the responding patients, and they had higher numbers of adaptive NK cells, CMV seropositivity, and a costimulatory cytokine environment before initiation of the treatment. Our results provide an understanding of the effects of anti–LAG-3+anti–PD-1 combination treatment in vivo in patients and highlight previously overlooked subpopulations of cells as targets of immune checkpoint therapies.

Single-cell profiling of anti-LAG3+anti–PD-1 treatment in patients with melFigure 1

Single-cell profiling of anti-LAG3+anti–PD-1 treatment in patients with melanoma. Schematic of the study cohorts and main findings. diff, difference; exp, expansion.

Results

Adaptive NK cells and CD8+ T cells have the highest LAG3 expression and are more numerous in responders. In total, we had 40 patients, 11 of whom were IO naive (7 of 11 [63.6%] were complete responders [CRs] or partial responders [PRs] and 4 [36.4%] had progressive disease [PD]) and 29 of whom were IO refractory (15 of 29 [51.7%] were CR/PR or had stable disease [SD] and 14 of 29 [48.3%] had PD). All the patients in the IO-naive cohort received 80+240 mg doses of relatlimab+nivolumab, while in the IO-refractory cohort 20 of 29 (68.9%) received 80+240 mg doses and 9 of 29 (31.0%) received 160+480 mg doses. The prior IO-refractory patients received previously anti–PD-1 therapy (22 of 29 [75.9%]) or anti–CTLA-4 and then anti–PD-1 (7 of 29 [24.1%]) (Table 1 and Supplemental Table 1).

With scRNA+TCRαβ-Seq, we profiled 18 peripheral blood (PB) samples from 5 IO-naive and 1 IO-refractory patients with metastatic melanoma treated with anti–LAG-3+anti–PD-1 combination therapy (CRs n = 2, PRs n = 1, PD n = 3; patient details are provided in Supplemental Table 1). We identified 24 cell clusters in the scRNA-Seq data (Figure 2, A and B), all of which were present in every sample, but their abundances varied between patients and time points (Supplemental Figure 1, A–E).

LAG3 is expressed at high levels in Tregs and CMV-associated adaptive NK ceFigure 2

LAG3 is expressed at high levels in Tregs and CMV-associated adaptive NK cells. (A) UMAP representation of CD45+-sorted cells in 18 scRNA+TCRαβ-Seq samples from 6 patients with melanoma before and after 4 weeks and 12 weeks of anti–LAG-3+anti–PD-1 treatment, profiled with scRNA+TCRαβ-Seq. (B) Scaled expression (expr) of selected differentially expressed markers (Padj < 0.05, Bonferroni-corrected t test) used to annotate clusters. The top row shows the log2 fold change (log2fc) of population abundances between patients with (CR/PR, n = 3) and without (PD, n = 3) a response at baseline. CM, central memory; Co-stim, costimulation; Co-inh, coinhibition; EM, effector memory; Mono, monocyte. (C) LAG3 expression at baseline as scaled, log2(× + 1) transformed values. The adjusted P value (Bonferroni-corrected t test) indicates the difference between adaptive NK cells and the other cell types. exh, exhausted. (D) scRNA-Seq population abundances between patients with (CR/PR, n = 3) and without (PD, n = 3) a response at baseline. P values were calculated with a Fisher’s 2-sided exact test, and significant values needed to have at least a |log2 fold change| >1. ***P < 0.001. (E) Proportion of CD56dimNKG2C+ adaptive NK cells among NK cells in IO-naive (CR/PR n = 7; SD/PD n = 4) and IO-refractory (CR/PR n = 3; SD/PD n = 26) groups at baseline. P values were calculated with the 2-sided Mann-Whitney U test. (F) Focused UMAP of NK cells, where the superimposed line corresponds to the predicted pseudotime maturation trajectory and scaled expression of markers used to identify the subpopulations. max, maximum; min, minimum. (G) UMAP representation of cells from 131 scRNA-Seq tumor biopsies or bone marrow aspirate samples from 10 different cancers profiled with 10× technology. Annotation was done with SingleR. (H) Proportion of LAG3+ cells across different cancers. P values were calculated with the Kruskal-Wallis test.

Prior to anti–LAG-3+anti–PD-1 treatment, we found that LAG3 was highly expressed in CD8+ T cells, CD4+ Tregs, and B cells, but the highest expression of LAG3 was surprisingly detected in adaptive NK cells in the scRNA-Seq data (adjusted P value [Padj] < 0.0001, Bonferroni-corrected t test, Figure 2C), which was validated by flow cytometry (n = 8, Supplemental Figure 2, A and B). The largest difference in cell population abundances between patients with a response (CR/PR) and without a response (PD) was seen in adaptive NK cells in the scRNA-Seq data (n = 3 vs. n = 3, P < 0.0001, Fisher’s 2-sided exact test, Figure 2, B and D). This analysis was extended with flow cytometric data, in which we saw a similar, albeit not statistically significant, trend (CD56dim

留言 (0)

沒有登入
gif