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. 2025 Sep 2;31(17):3702-3714.
doi: 10.1158/1078-0432.CCR-24-2499.

Unraveling Relatlimab-Specific Biology Using Biomarker Analyses in Patients with Advanced Melanoma in RELATIVITY-047

Evan J Lipson  1 , Sonia Dolfi  2 , Hao Tang  2 , Hussein A Tawbi  3 , Francisco Medina-Soto  4 , Erika Castillo Gutiérrez  5 , Piotr Rutkowski  6 , Helen Gogas  7 , Emilio Murillo  8 , Paolo A Ascierto  9 , Keyur Desai  2 , Michele Maio  10 , Korey Demers  2 , Antonella Mazzei  2 , Sarah Keidel  2 , Karen Miller-Moslin  2 , Jia Xin Yu  2 , F Stephen Hodi  11 , Dirk Schadendorf  12 , Georgina V Long  13 , Charlie Garnett-Benson  2
Affiliations

Unraveling Relatlimab-Specific Biology Using Biomarker Analyses in Patients with Advanced Melanoma in RELATIVITY-047

Evan J Lipson et al. Clin Cancer Res. .

Abstract

Purpose: Administration of the lymphocyte activation gene 3 (LAG-3) inhibitor relatlimab (RELA) and the PD-1 inhibitor nivolumab (NIVO) significantly prolonged progression-free survival (PFS) versus NIVO alone in patients with advanced melanoma treated in RELATIVITY-047. This report describes correlative analyses of biospecimens collected within that trial to better understand the mechanisms of action and identify patients who could benefit from treatment with NIVO + RELA.

Patients and methods: Pre- and on-treatment peripheral blood samples from 563 patients were analyzed using flow cytometry for changes in 77 prespecified immune cell populations and using immunoassay for peripheral IFNγ. Pretreatment tumor biopsies were evaluated using IHC and RNA sequencing.

Results: On-treatment expansion of 25 peripheral immune cell populations was significantly greater with NIVO + RELA versus NIVO alone. Responders to NIVO + RELA had greater on-treatment increases in LAG-3+CD4+ T cells than nonresponders. Significantly greater increases in peripheral IFNγ occurred after treatment with NIVO + RELA versus NIVO alone. A longer PFS was observed in patients treated with NIVO + RELA whose tumors had low CD8 expression compared with the NIVO arm. When evaluating the co-expression of CD8 and LAG-3, patients whose tumors had high CD8+LAG-3+ also showed a PFS benefit with NIVO + RELA versus NIVO. RNA sequencing revealed several distinct gene signatures associated with response to NIVO + RELA.

Conclusions: These results highlight the unique biological effects of RELA in combination with NIVO and provide further understanding of the patient characteristics associated with increased benefit from NIVO + RELA.

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Conflict of interest statement

E.J. Lipson reports grants from Bristol Myers Squibb during the conduct of the study; grants and personal fees from Bristol Myers Squibb, Merck, Regeneron Pharmaceuticals, and Sanofi; and personal fees from CareDx, Genentech, Instil Bio, Natera, Nektar Therapeutics, Pfizer, Replimune, Sun Pharma, IO Biotech, HUYA Bioscience, Novartis, OncoSec, and Merck outside the submitted work; and stock ownership (<$10K) in Iovance Biotherapeutics. S. Dolfi is an employee of Bristol Myers Squibb with stock benefits. H. Tang reports personal fees from Bristol Myers Squibb during the conduct of the study. H.A. Tawbi reports personal fees from Bristol Myers Squibb, Corcept Therapeutics, IO Biotech, Immunocore Holdings, Iovance Biotherapeutics, Krystal Biotech, Medicenna, Pfizer, Strand Therapeutics, and T-knife Therapeutics; grants and personal fees from Eisai, Merck, Novartis, and Regeneron Pharmaceuticals; and grants from Dragonfly Therapeutics, Genentech, GSK, and Syntrix Pharmaceuticals outside the submitted work. P. Rutkowski reports personal fees from MSD, Bristol Myers Squibb, Pierre Fabre, Medison Pharma, GENESIS Pharma, and Novartis outside the submitted work. H. Gogas reports grants from Bristol Myers Squibb during the conduct of the study; personal fees from MSD, Regeneron Pharmaceuticals, Pierre Fabre, and Pfizer; and grants from GENESIS Pharma, Pfizer, and Pierre Fabre outside the submitted work. P.A. Ascierto reports grants and personal fees from Bristol Myers Squibb, Roche-Genentech, Regeneron Pharmaceuticals, and Pfizer; personal fees and other support from MSD, Pierre Fabre, BioAI Health, Replimune, and Philogen; and personal fees from Novartis, Sun Pharma, Immunocore Holdings, Italfarmaco, Boehringer Ingelheim, Nouscom, Medicenna, Valo Therapeutics, Bayer Healthcare Pharmaceuticals, Erasca, BioNTech, ANAVEON, Genmab, Menarini, and Incyte outside the submitted work. K. Desai reports other support from Bristol Myers Squibb outside the submitted work. M. Maio reports other support from Bristol Myers Squibb, Roche, MSD, Merck Serono, Eli Lilly and Company, Amgen, Novartis, GSK, SciClone Pharmaceuticals, iOnctura, Regeneron Pharmaceuticals, and Astex Pharmaceuticals during the conduct of the study, as well as other support from Bristol Myers Squibb, Roche, Merck Serono, MSD, Eli Lilly and Company, Amgen, Novartis, GSK, Regeneron Pharmaceuticals, Pierre Fabre, iOnctura, SciClone Pharmaceuticals, and Astex Pharmaceuticals outside the submitted work. A. Mazzei reports personal fees from Bristol Myers Squibb during the conduct of the study. S. Keidel reports other support from Bristol Myers Squibb during the conduct of the study, as well as other support from Moderna and AbbVie outside the submitted work. K. Miller-Moslin reports other support from Bristol Myers Squibb during the conduct of the study, as well as other support from Bristol Myers Squibb outside the submitted work. J.X. Yu reports employment with Bristol Myers Squibb and stock benefits as an employee. F.S. Hodi reports nonfinancial support and other support from Bristol Myers Squibb during the conduct of the study; grants and personal fees from Bristol Myers Squibb; and personal fees from Merck, Novartis, Compass Therapeutics, Apricity Health, Bicara Therapeutics, Checkpoint Therapeutics, BioEntre Therapeutics, Gossamer Bio, Iovance Biotherapeutics, CatalYm, Immunocore Holdings, Kairos Pharma, Rheos Medicines, Bayer Healthcare Pharmaceuticals, Zumutor Biologics, Corner Therapeutics, Puretech Health, Curis Lifesciences, AstraZeneca, Pliant Therapeutics, Solu Therapeutics, Vir Biotechnology, and 92Bio outside the submitted work; and a patent for Methods for Treating MICA-related Disorders (No. 20100111973) pending, a patent for Tumor Antigens and Uses Thereof (No. 7250291) issued, a patent for Angiopoietin-2 Biomarkers Predictive of Anti-immune Checkpoint Response (No, 20170248603) pending, a patent for Compositions and Methods for Identification, Assessment, Prevention, and Treatment of Melanoma using PD-L1 Isoforms (No, 20160340407) pending, a patent for Therapeutic Peptides (No, 20160046716) pending, a patent for Methods of Using Pembrolizumab and Trebananib pending, a patent for Vaccine Compositions and Methods for Restoring NKG2D Pathway Function against Cancers (No. 10279021) issued, a patent for Antibodies That Bind to MHC Class I Polypeptide-related Sequence A (No. 10106611) issued, a patent for Anti-galectin Antibody Biomarkers Predictive of Anti-immune Checkpoint and Anti-angiogenesis Responses (publication number: 20170343552) pending, and a patent for Antibodies against EDIL3 and Methods of Use Thereof pending. D. Schadendorf reports grants from Merck and personal fees from Merck, Bristol Myers Squibb, Pierre Fabre, AstraZeneca, Boehringer Ingelheim, Ipsen, Sun Pharma, Philogen, Replimune, BioAlta, Daiichi Sankyo, Formycon, BioNTech, Immatics, Immunocore Holdings, NeraCare, Merck Serono, Novartis, Regeneron Pharmaceuticals, Iovance Biotherapeutics, and SkylineDX outside the submitted work. G.V. Long reports personal fees from Agenus, Amgen, Array BioPharma, AstraZeneca, Bayer Healthcare Pharmaceuticals, BioNTech, Boehringer Ingelheim, Bristol Myers Squibb, Evaxion Biotech, Fortiva Biologics, GI Innovation, Hexal AG, Highlight Therapeutics, Immunocore Holdings, Innovent Biologics USA, IO Biotech, Iovance Biotherapeutics, MSD, Novartis, PHMR Limited, Pierre Fabre, Regeneron Pharmaceuticals, Scancell, and SkylineDX outside the submitted work. C. Garnett-Benson is an employee of Bristol Myers Squibb with stock benefits. No disclosures were reported by the other authors.

Figures

Figure 1.
Figure 1.
Increases in peripheral Ki67+ CM and EM T-cell subsets. Horizontal lines within each graph represent the median biomarker level in each arm at each time point. P values were estimated using the paired Wilcoxon test from baseline and 4-week (day 29) paired samples.
Figure 2.
Figure 2.
Changes in peripheral immune cell subsets in the NIVO + RELA arm compared with the NIVO arm. A, Degree of on-treatment change in T-cell subsets within the NIVO + RELA (n = 236) and NIVO (n = 231) arms. Each line represents an individual cell population and is labeled with the corresponding cell population listed in (B), which shows the change from baseline to 4 weeks (day 29) in T-cell subsets. Darker blue, NIVO larger fold change; darker red, NIVO + RELA larger fold change. aNominal P values using the paired Wilcoxon test were not adjusted for multiplicity. bAll patients, paired only. Relative difference is the difference between arms in fold changes of cell populations, whereas relative difference % is the difference between arms in fold changes as a percentage of the observed frequency in the NIVO arm. cLAG-3, cytoplasmic/intracellular LAG-3; FOXP3, forkhead box P3; HLA-DR, human leukocyte antigen, DR isotype; sLAG-3, surface LAG-3.
Figure 3.
Figure 3.
Significant changes in additional immune cell subsets within the NIVO + RELA and NIVO arms. A, Degree of on-treatment change across select cell populations within the NIVO + RELA (n = 236) and NIVO (n = 231) arms. Each line represents an individual cell population and is labeled with the corresponding cell population listed in (B), which shows the change from baseline to 4 weeks (day 29) in select cell populations. Delta is the difference in observed frequency (%) between baseline and 4 weeks. Darker blue, a lower delta/% (fold) change; darker red, a higher delta/% (fold) change. cLAG-3, cytoplasmic/intracellular LAG-3; FOXP3, forkhead box P3; HLA-DR, human leukocyte antigen, DR isotype; ns, not significant; sLAG-3, surface LAG-3.
Figure 4.
Figure 4.
On-treatment fold changes by best overall response in LAG-3+CD4+ T cells in patients treated with NIVO + RELA or NIVO. Samples were collected after treatment and at week 4 on therapy. aDefined as CD45+, CD3+, CD56−, CD4+, CD8−, intracellular LAG-3+, sLAG-3+ cells as a percentage of CD45+, CD3+, CD56−, CD4+, CD8− cells. P values were adjusted for multiple testing (Bonferroni correction). Samples with an undetectable cell population at baseline were excluded. sLAG-3, surface lymphocyte activation gene-3.
Figure 5.
Figure 5.
Serum IFNγ changes after treatment with NIVO + RELA and NIVO alone. A, Relative change in serum IFNγ levels at baseline, 4 weeks, and 8 weeks after treatment initiation. B, Fold changes in serum IFNγ levels and association with best overall response. Error bars indicate 95% CI.
Figure 6.
Figure 6.
Association of CD8 expression with PFS in patients treated with NIVO + RELA or NIVO. A, PFS by CD8 tertile within treatment arms. B, Between-arm comparison by CD8 tertile. CD8 tertile (%) definitions: CD8 low = (0, 3.52); CD8 medium = (3.52, 11.93); CD8 high = (11.93, 56.05). aAssay: CD8 monoplex IHC. bSamples with complete CD8, LAG-3, and PD-L1 by monoplex IHC (n = 474). mPFS, median PFS; NA, not applicable.
Figure 7.
Figure 7.
Association of LAG−3+/−CD8+ tumors with PFS in patients treated with NIVO + RELA compared with NIVO. A, Association of LAG-3−CD8+ and (B) LAG-3+CD8+ tumors with PFS by tertile (high) between treatment arms. Assay: CD8/LAG-3 multiplex IHC HALO analysis. mPFS, median PFS.
See this image and copyright information in PMC

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