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622 Immune phenotype and iFRET functional analysis are biomarkers of response to neoadjuvant intralesional therapy for high risk stage II melanoma
  1. Amanda Kirane1,
  2. Michael C Lowe2,
  3. Mohammad Usman Ahmad1,
  4. Saurabh Sharma3,
  5. Mamatha Serasanambati1,
  6. Allison B Warner4,
  7. Chris Applebee5,
  8. Elena Safrygina5,
  9. Susan Swetter1,
  10. Sunil Reddy6,
  11. Emanual Maverakis7 and
  12. Banafshe Larijani5
  1. 1Stanford University, Palo Alto, CA, USA
  2. 2Emory University School of Medicine, Atlanta, GA, USA
  3. 3School of Medicine, Stanford University, Stanford, CA, USA
  4. 4Memorial Sloan Kettering Cancer Center, New York, NY, USA
  5. 5University of Bath, Bath, UK, UK
  6. 6Stanford Cancer Institute, Redwood City, CA, USA
  7. 7University of California, Davis, Sacramento, CA, USA
  • Journal for ImmunoTherapy of Cancer (JITC) preprint. The copyright holder for this preprint are the authors/funders, who have granted JITC permission to display the preprint. All rights reserved. No reuse allowed without permission.


Background Although survival remains significantly poorer for thick, ulcerated melanoma, application of immunotherapy is controversial in the absence of nodal disease. Talimogene Laherparpvec (TVEC) is the only FDA approved intralesional therapy for melanoma and has demonstrated overall response and bystander effect (34% complete response in uninjected lesions) in late stage melanoma. We hypothesize neoadjuvant TVEC in stage II melanoma will treat local and subclinical regional disease and still allow standard of care surgery.

Methods We present an open-label, Phase 2 pilot study (NCT04427306) of TVEC in stage II melanoma patients with residual disease prior to wide excision and sentinel lymph node biopsy (SLNB). Primary endpoints were safety and efficacy of TVEC with secondary objective to identify of immunologic and molecular predictors of response by immuneFRET (iFRET), digital spatial profiling, serum luminex, and immune-typing of PBMCs.

Results To date, 6 patients were enrolled and treated. All patients completed therapy, adverse events were limited to grade I, there were no surgical complications. 3 patients had at least partial pathologic response (<50% viable tumor), 1 complete response, and 1 positive for sentinel disease. No patients experienced relapse or conversion to systemic therapy at follow up. iFRET revealed dynamic shifts in functional PD-1 receptor engagement with increases correlating to positive therapeutic response and serum pro-inflammatory profile and decreases reflected immunosuppressive profiles. PD-1 expression alone did not correlate to response. Cytof profiling revealed common trends in T cell population shifts (CD3, CD4, CD8) that did not correlate to therapeutic response, but significant divergences in monocyte subsets (alternatively vs classically activated), regulatory cells (Treg, NKreg, MDSC), T cell activation, and migratory T cell subsets (figure 1).

Conclusions This study is the first to neoadjuvantly target earlier-stage melanoma with intra-tumoral immunotherapy and mechanistically dissect immune factors responsible for response and bystander effect in draining nodes. Neoadjuvant TVEC was safe and well tolerated. Baseline characteristics and early functional shifts may differ between patients likely to benefit from oncolytic virus versus checkpoint blockade. We expect the granularity of our data to contribute significantly to future studies of precision application of immunotherapy in high risk patients.

Ethics Approval Ethics approval was obtained via IRB committee (IRB 68118). All participants provided informed consent for study participation.

Consent Written informed consent was obtained from the patient for publication of this abstract and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See

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