Several high-profile publications and presentations demonstrate the value of Rules-Based Medicine’s MAP (Multi-Analyte Profile) and Simoa platforms for measuring panels of immune- and cancer-related soluble proteins present in peripheral blood from patients enrolled in immuno-oncology clinical trials.

IFN-gamma chemokines CXCL9, CXCL10, IL-2RA were increased with budigalimab treatment, consistent with other PD-1 inhibitors. 1,2,3

1. Model informed dosing regimen and phase I results of the anti-PD-1 antibody budigalimab (ABBV-181). (2020) Powderly J, et al. Clin Trans Sci. 2020 Aug 8. doi: 10.1111/cts.12855.

2. Safety, pharmacokinetics, and efficacy of budigalimab with rovalpituzumab tesirine in patients with small cell lung cancer. (2021) Calvo E, et al. Cancer Treatment and Research Communications. Volume 28, 2021, 100404. org/10.1016/j.ctarc.2021.100405

3. First-in-human phase 1 study of budigalimab, an anti-PD-1 inhibitor, in patients with non-small cell lung cancer and head and neck squamous cell carcinoma. (2020) Italiano A, et al. Cancer Immunology, Immunotherapy. org/10.1007/s00262-021-02973-w

Serum inflammatory biomarkers (IFN-gamma, IP-10, IL-18) showed does schedule-specific effects immune signatures with same-day dosing of Brentuximab vedotin in combination with Nivolumab compared to staggered dosing.4

4. Brentuximab vedoin in combination with nivolumab in relapsed or refractory Hodgkin lymphoma: 3-year study results. (2021) Advani RH, et al. Blood. 2021 Aug 12;138(6):427-438. doi: 10.1182/blood.2020009178.

CXCR4 inhibitor mavorixafor + nivolumab show increased CXCL9 levels from baseline corresponding to antitumor activity in RCC patients who did not respond to nivolumab monotherapy.5

5. A phase 1b trial of the CXCR4 inhibitor mavorixafor and nivolumab in advanced renal cell carcinoma patients with no prior response to nivolumab monotherapy. (2020) Choueiri TK, et al. Invet New Drugs. doi.org/10.1007/s10637-020-01058-2

CSF-1R inhibitor LY3022855 increased serum CSF-1 and IL-34 levels suggesting CSF-1R target engagement and apoptosis of tumor associated macrophages.6

6. LY3022855, an anti-colony stimulating factor-1 receptor (CSF-1R) monoclonal antibody, in patients with advanced solid tumors refractory to standard therapy: phase 1 dose-escalation trial. (2021) Dowalti A, et al. Invest New Drugs. doi: 10.1007/s10637-021-01084-8.

Serum immune markers at baseline and on-therapy are associated with survival and tumor regression with checkpoint inhibition therapies. 7

7. Serum interleukin-6 and C-reactive protein are associated with survival in melanoma patients receiving immune checkpoint inhibition. (2020) Laino AS, et al. J Immunother Cancer. 2020;8:e000842. doi:10.1136/jitc-2020-000842

In patients with chronic lymphocytic leukemia, median increases in inflammatory cytokine levels were lower with ibrutinib-obinutuzumab treatment versus chlorambucil-obinutuzumab.8

8. Pretreatment with ibrutinib reduces cytokine secretion and limits the risk of obinutuzumab-induced infusion-related reactions in patients with CLL: analysis from the iLLUMINATE study. (2021) Greil R, et al. Ann Hematol. 2021 Jul;100(7):1733-1742. doi: 10.1007/s00277-021-04536-6. Epub 2021 May 20.

Baseline serum biomarker signatures prognostic for nivolumab drug clearance & overall survival in melanoma and renal cancer. 9,10

9. Development of a prognostic composite cytokine signature based on the correlation with nivolumab clearance: translational PK/PD analysis in patients with renal cell carcinoma. (2019) Wang R, et al. J Immunother Cancer. 2019 Dec 11;7(1):348. doi: 10.1186/s40425-019-0819-2.

10. A machine-learning approach to Identify a prognostic cytokine signature that is associated with nivolumab clearance in patients with advanced melanoma. (2019) Wang R, et al. Clin Pharmacol Ther. 2020 Apr;107(4):978-987. doi: 10.1002/cpt.1724.

Circulating immune and T-cell activation biomarker levels elevated in CX-072 treatment are consistent with PD-1/PD-L1 pathway activation.11

11. Evidence of intratumoral localization, activation, and immunomodulatory effect of CX-072, a PROBODY therapeutic targeting PD-L1, in a phase ½ trial. (2020) Lyman SK, et al. P310 ASCO20 Virtual Scientific Program May 29-31.

Elevated baseline levels of circulating inflammatory, myeloid (CRP, IL-6, S100A12) and angiogenic proteins (ANGPT2, vWF, ICAM1) correlated with shorter OS.12

  1. Correlation of angiogenic and immunomodulatory proteins with clinical outcomes of durvalumab (anti-PD-L1) in recurrent/metastatic head and neck squamous cell carcinoma. (2019) Guo X, et al. P6048 ASCO June 1-4. Chicago, IL.

High circulating levels of IL-6 is correlated with poor prognosis of HCC patients and positively correlated with tumor PD-L1 expression.13

  1. IL-6/JAK1 pathway drives PD-L1 Y112 phosphorylation to promote cancer immune evasion. (2019) Chan LC, et al. J Clin Invest. 2019 Jul 15;129(8):3324-3338. doi: 10.1172/JCI126022.

TGFb inhibition: immune-related biomarkers with pharmacodynamic, prognostic and predictive potential.14, 15,16,17

  1. TGFb receptor inhibitor galunisertib is linked to inflammation- and remodeling-related proteins in patients with pancreatic cancer. (2019) Melisi D, et al. Cancer Chemother Pharmacol. 2019 May; 83(5):975-991. doi: 10.1007/s00280-019-03807-4.
  2. Phase 1b/2a study of galunisertib, a small molecule inhibitor of transforming growth factor-beta receptor I, in combination with standard temozolomide-based radiochemotherapy in patients with newly diagnosed malignant glioma. (2020) Wick A, et al. Invest New Drugs. 2020 Oct;38(5):1570-1579. doi: 10.1007/s10637-020-00910-9.
  3. Biomarkers and overall survival in patients with advanced hepatocellular carcinoma treated with TGF-beta R1 inhibitor galunisertib. (2020) Giannelli G, et al. PLoS ONE. 2020 Mar 25;15(3):e0222259. doi: 10.1371/journal.pone.0222259.
  4. AVID200, first-in-class TGF-beta 1 and 3 selective and potent inhibitor: safety and biomarker results of a phase I monotherapy dose-escalation study in patients with advanced solid tumors. (2020) Yap TA, et al. P3587. ASCO Virtual Scientific Program. 2020. https://meetinglibrary.asco.org/record/187808/abstract

TruCulture cytokine analysis of adenosine A2A receptor engagement and dose dependent pharmacodynamic release of immune suppression.18

  1. Immune checkpoint inhibition in sepsis: a phase 1b randomized study to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamics of nivolumab. (2019) Hotchkiss RS, et al. Intensive Care Med. 2019 Oct;45(10) 1360-1371.

Circulating levels of IFNγ as measured by an ultrasensitive immunoassay are pharmacodynamic for an immune stimulating agent and prognostic for OS.16

18. EOS100850 inhibits A2A receptor signaling in human whole blood: two pharmacodynamic assays to monitor EOS100850 activity in clinical studies. (2019) Martinoli C, et al. P4154. AACR Mar 29- Apr 3. Atlanta, GA.

Elevated levels of serum inflammatory proteins including APO E pre-vaccination are associated with the clinical non-responder phenotype and shorter PFS.19

19. Serum apolipoprotein E and other inflammatory markers can identify non-responding patients to a dendritic cell vaccine. (2019) Leeman H, et al. Transl Oncol. 2019 Mar;12(3):397-403. doi: 10.1016/j.tranon.2018.11.002.

Activation of the immune system in a complete responder as reflected by levels of peripheral interferon-γ-related cytokines (IL-18, CXCL10), increased proliferating NK and CD8+ T cells, and tumor immune microenvironment signatures.20

20. Tumor immune microenvironment and genomic evolution in a patient with metastatic triple negative breast cancer and a complete response to atezolizumab. (2019) Molinero L, et al. J Immunother Cancer. 2019 Oct 23;7(1):274. doi: 10.1186/s40425-019-0740-8.

A single nivolumab dose does not cause unexpected safety findings or ‘cytokine storm’ in sepsis patients with organ dysfunction and low lymphocyte counts.21

21. Immune checkpoint inhibition in sepsis: a phase 1b randomized study to evaluate the safety, tolerability, pharmacokinetics and pharmacodynamics of nivolumab. (2019) Hotchkiss RS, et al. Intensive Care Med. 2019 Oct;45(10) 1360-1371. doi: 10.1007/s00134-019-05704-z.

Circulating levels of IFNγ as measured by an ultrasensitive immunoassay are pharmacodynamic for an immune therapies.22,23

22. Combination of atezolizumab and obinutuzumab in patients with relapsed-refractory follicular lymphoma and diffuse large B-cell lymphoma: Results from a phase 1 study. (2021) Palomba ML, et al. Clinical Lymphoma Myeloma and Leukemia. ISSN 2152-2650 org/10.1016/j.clml.2021.12.010.

23. The immunomodulatory effects of cancer therapy on IFN-gamma responses in the periphery. (2018) Brunet LA, et al. P81 SITC. Nov 9-11. National Harbor, MD.

Circulating protein biomarkers including soluble Axl are pharmacodynamic & potentially predicative of patient response.24,25,26,27

24. Durable responses observed in elderly AML patients unfit for intensive chemotherapy with first-in-class selective AXL inhibitor bemcentinib (BGB324) in combination with LDAC: phase II open-label study. (2019) Loges S, et al. P3943. ASH, Dec 9. Orlando, FL.

25. Phase II open-label, multi-centre study of bemcentinib (BGB324), a first-in-class selective AXL inhibitor, in combination with pembrolizumab in patients with advanced NSCLC. (2018) Lorens J, et al. P3078. ASCO June 1-4. Chicago, IL.

26. Analysis of anti-leukemic activity, predictive biomarker candidates, immune activation and pharmacodynamics in R/R AML and MDS in response to treatment with bemcentinib (BGB324), a first-in-class selective AXL inhibitor, in a phase II open label, multi-centre study. (2018) Gjertsen BT, et al. P7020. ASCO June 1-4. Chicago, IL.

27. A randomized phase Ib/II study of the selective small molecule AXL inhibitor bemcentinib (BGB324) in combination with either dabrafenib/trametinib or pembrolizumab in patients with metastatic melanoma. (2018) Oddbjorn S, et al. P3078. ASCO June 1-4. Chicago, IL.

A large cross-sectional retrospective study of over 2,000 patients demonstrating that serum IL-8 is a promising clinical biomarker for patients that benefit from I-O therapy.28,29

28. Elevated serum interleukin-8 is associated with enhanced intratumor neutrophils and reduced clinical benefit of immune-checkpoint inhibitors. (2020) Schalper KA, et al. Nat Med. 2020 May;26(5):688-692. doi: 10.1038/s41591-020-0856-x.

29. Serum interleukin 8 (IL-8) may serve as a biomarker of response to immuno-oncology (I-O) therapy. (2018) Carleton M, et al. P3025. ASCO June 1-4. Chicago, IL.

If high levels of IL-8 reduces the effectiveness of anti-PD-1 treatment, neutralization of IL-8 in combination with checkpoint blockade is an attractive combination therapy approach.30

30. Phase I trial of BMS-986253, an anti-IL-8 monoclonal antibody, in patients with metastatic or unresectable solid tumors. (2018) Collins J, et al. P3091. ASCO June 1-4. Chicago, IL.

 Serum immune markers increased from baseline with combination lenvatinib and pembrolizumab treatment in patients who achieved response.31

31. Biomarker results and preclinical rationale for combination lenvatinib and pembrolizumab in advanced endometrial carcinoma. (2018) Makker V, et al. P5597. ASCO June 1-5. Chicago, IL.

IL-8 biomarker guided precision medicine phase1/2b immuno-oncology trial.32

32. Phase 1b/2 study of nivolumab in combination with an anti–IL-8 monoclonal antibody, BMS-986253, in a biomarker enriched population of patients with advanced cancer. (2018) Melero I, et al. P TPS3109. ASCO June 1-4. Chicago, IL.

OX-40 agonist in combination with NIVO or IPI increases serum IFN-g and IP-10 (CXCL10) levels.33

33. From bench to bedside: exploring OX40 receptor modulation in a phase 1/2a study of the OX40 costimulatory agonist BMS-986178 ± nivolumab (NIVO) or ipilimumab (IPI) in patients with advanced solid tumors. (2018) Wang R, et al. LB-127. AACR April 14-18. Chicago, IL.

PEGylated IL-10 (Pegilodecakin) induces sustained elevation of Th1 and Th2 cytokines in serum.34,35,36

34. PEGylated IL-10 (Pegilodecakin) induces systemic immune activation, CD8+ T cell invigoration and polyclonal T cell expansion in cancer patients. (2018) Naing A, et al. Cancer Cell. 2018 Nov 12;34(5):775-791.e3. doi: 10.1016/j.ccell.2018.10.007.

35. Safety, antitumor activity, and immune activation of pegylated recombinant human interleukin-10 (Am0010) in patients with advanced solid tumors. (2016) Naing A, et al. J Clin Oncol. 2016 Oct 10;34(29):3562-35569. doi: 10.1200/JCO.2016.68.1106.

36. Phase I study with PEGylated human IL-10 (AM0010) alone or in combination with anti-PD-1 or FOLFOX : immune biomarker update. (2017) Naing A, et al. SITC Nov 9-11. National Harbor, MD.

Four plasma proteins correlate with clinical benefit of Axl kinase inhibition that reverses suppression of immune responses in AML patients. 37

37. The orally available selective AXL inhibitor BGB324 (bemcentinib) induces diversification of the immune repertoire and specific changes in plasma biomarker profiles. (2017) Loges S, et al. ASH, Dec 9-12. Atlanta, GA.

Low baseline myeloid cell-associated serum proteins (M-CSF, IL-8, and IL-6) and high baseline IFNγ-inducible proteins (CXCL9,CXCL10, and TWEAK) were associated with longer OS.38

38. Differential association of myeloid cell and IFN-γ associated proteins with clinical response to durvalumab treatment in urothelial bladder cancer. (2017) Guo X, et al. P32. SITC Nov 8-12. National Harbor, MD.

TruCulture immunoprofiling of antigen-stimulated blood has a variety of applications in I-/O clinical trials.39,40,41

39. Preclinical investigations and a first-in-human phase I trial of M4112, the first dual inhibitor of indoleamine 2,3-dioxygenase 1 and tryptophan 2,3-dioxygenase 2, in patients with advanced solid tumors. Naing A, et al. (2020) J ImmunoTher Cancer 2020;8:e000870. doi:10.1136/jitc-2020-000870

40. Results of the phase I open label clinical trial SAKK 06/14 assessing safety of intravesical instillation of VPM1002BC, a recombinant mycobacterium Bacillus Calmette Guerin (BCG), in patients with non-muscle invasive bladder cancer and previous failure of conventional BCG therapy. (2020) Rentsch CA, et al. OncoImmunology. 2020 Apr 21;9:1, 1748981, doi:10.1080/2162402X.2020.1748981.

41. Identifying baseline immune-related biomarkers to predict clinical outcome of immunotherapy. (2017) Gnjatic S, et al. J Immunother Cancer. 2017 May 16;5:44. doi: 10.1186/s40425-017-0243-4.

Low baseline serum IFNγ is associated with improved OS upon subsequent immunotherapy with bavituximab + docetaxel treatment.42

42. IFNγ analysis in blood and tissue as a potential prognostic and/or predictive biomarker. (2017) Kallinteris NL, et al. P458. AACR April 1-5, Washington D.C.

Serum biomarker changes from baseline reflect nivolumab pharmacodynamics in the tumor microenvironment.43,44

43. Molecular correlates of response to nivolumab at baseline and on treatment in patients with RCC. (2020) Ross-Macdonald P, et al. J Immunother Cancer. 2021 Mar;9(3):e001506. https://pubmed.ncbi.nlm.nih.gov/33658305/doi: 10.1136/jitc-2020-001506.

44. Immunomodulatory activity of nivolumab in metastatic renal cell carcinoma. (2015) Choueiri TK, et al. Clin Cancer Res. 2016 Nov 15;22(22):5461-5471. doi: 10.1158/1078-0432.CCR-15-2839.

Dose-dependent urelumab exposures were associated with peripheral NK and T-cell activation and an increase in IFNγ-induced serum cytokines (CXCL9, CXCL10, MIP-1β, MCP-2). 45,46

45. Urelumab alone or in combination with rituximab in patients with relapsed refractory B-cell lymphoma. (2020) Timmerman J, et al. Am J Hematol. 2020 May;95(5):510-520. doi: 10.1002/ajh.25757.

46. Assessing the potential for enhanced antibody-dependent cell-mediated cytotoxicity by combining the CD137 antibody urelumab with rituximab or cetuximab in patients with refractory lymphoma or select advanced solid tumors. (2016) Segal NH, et al. P267. SITC Nov 9-13. National Harbor, MD.

Dose-proportional increase in serum sIL-2Ra and IP-10 (CXCL10) with anti–LAG-3 + nivolumab treatment.47

47. Initial experience administering BMS-986016, a monoclonal antibody that targets lymphocyte activation gene (LAG)–3, alone and in combination with nivolumab to patients with hematologic and solid malignancies. (2016) Evan JL, et al. P238. SITC Nov 9-13. National Harbor, MD.