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Preclinical Evaluation of LVR01 Attenuated Salmonella as Neoadjuvant Intralesional Therapy in Combination with Chemotherapy for Melanoma Treatment

Published:October 14, 2021DOI:
      Treatment of malignant melanoma has improved in the last few years owing to early detection and new therapeutic options. Still, management of advanced disease remains a challenge because it requires systemic treatment. In such cases, dacarbazine-based chemotherapy has been widely used, despite low efficacy. Neoadjuvant therapies emerge as alternative options that could help chemotherapy to achieve increased benefit. In this work, we evaluate LVR01, an attenuated Salmonella enterica serovar typhimurium, as neoadjuvant intralesional therapy in combination with dacarbazine in a preclinical melanoma model. B16F1 melanoma‒bearing mice received intraperitoneal administration of dacarbazine for 3 consecutive days. LVR01 treatment, consisting of one single intratumoral injection, was applied 1 day before chemotherapy began. This therapeutic approach retarded tumor growth and prolonged overall survival, revealing a strong synergistic antitumor effect. Dacarbazine induced a drastic reduction of secondary lymphoid organ cellularity, which was partially restored by Salmonella, particularly potentiating activated cytotoxic cell compartments. Systemic immune reactivation could be a consequence of the intense inflammatory tumor microenvironment induced by LVR01. We propose that the use of LVR01 as neoadjuvant intralesional therapy could be considered as an interesting strategy with close clinical application to boost chemotherapy effect in patients with melanoma.


      DTIC (dacarbazine), p.t.i. (post-tumor implantation), TDLN (tumor-draining lymph node)
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        • Agorio C.
        • Schreiber F.
        • Sheppard M.
        • Mastroeni P.
        • Fernandez M.
        • Martinez M.A.
        • et al.
        Live attenuated Salmonella as a vector for oral cytokine gene therapy in melanoma.
        J Gene Med. 2007; 9: 416-423
        • Andtbacka R.H.
        • Kaufman H.L.
        • Collichio F.
        • Amatruda T.
        • Senzer N.
        • Chesney J.
        • et al.
        Talimogene laherparepvec improves durable response rate in patients with advanced melanoma.
        J Clin Oncol. 2015; 33: 2780-2788
      1. Babjuk M, Burger M, Compérat E, Gontero P, Mostafid AH, Palou J, et al. EUA guidelines on non-muscle-invasive bladder cancer (TaT1 and CIS) [EAU guidelines]. Edn. Paper presented at: EAU Annual Congress. 17–21 July 2020; Amsterdam, The Netherlands.

        • Bascuas T.
        • Moreno M.
        • Grille S.
        • Chabalgoity J.A.
        Salmonella immunotherapy improves the outcome of CHOP chemotherapy in non-Hodgkin lymphoma-bearing mice.
        Front Immunol. 2018; 9: 7
        • Chabalgoity J.A.
        • Moreno M.
        • Carol H.
        • Dougan G.
        • Hormaeche C.E.
        Salmonella typhimurium as a basis for a live oral Echinococcus granulosus vaccine.
        Vaccine. 2000; 19: 460-469
        • Chapman P.B.
        • Hauschild A.
        • Robert C.
        • Haanen J.B.
        • Ascierto P.
        • Larkin J.
        • et al.
        Improved survival with vemurafenib in melanoma with BRAF V600E mutation.
        N Engl J Med. 2011; 364: 2507-2516
        • Cunningham C.
        • Nemunaitis J.
        A phase I trial of genetically modified Salmonella typhimurium expressing cytosine deaminase (TAPET-CD, VNP20029) administered by intratumoral injection in combination with 5-fluorocytosine for patients with advanced or metastatic cancer. Protocol no: CL-017. Version: April 9, 2001.
        Hum Gene Ther. 2001; 12: 1594-1596
        • Forbes N.S.
        • Coffin R.S.
        • Deng L.
        • Evgin L.
        • Fiering S.
        • Giacalone M.
        • et al.
        White paper on microbial anti-cancer therapy and prevention.
        J Immunother Cancer. 2018; 6: 78
        • Foucquier J.
        • Guedj M.
        Analysis of drug combinations: current methodological landscape [published correction appears in Pharmacol Res Perspect 2019;7:e00549].
        Pharmacol Res Perspect. 2015; 3: e00149
        • Fritzell S.
        • Sandén E.
        • Eberstål S.
        • Visse E.
        • Darabi A.
        • Siesjö P.
        Intratumoral temozolomide synergizes with immunotherapy in a T cell-dependent fashion.
        Cancer Immunol Immunother. 2013; 62: 1463-1474
        • Fujimura T.
        • Kakizaki A.
        • Kambayashi Y.
        • Sato Y.
        • Tanita K.
        • Lyu C.
        • et al.
        Cytotoxic antimelanoma drugs suppress the activation of M2 macrophages.
        Exp Dermatol. 2018; 27: 64-70
        • Grille S.
        • Moreno M.
        • Bascuas T.
        • Marqués J.M.
        • Muñoz N.
        • Lens D.
        • et al.
        Salmonella enterica serovar typhimurium immunotherapy for B-cell lymphoma induces broad anti-tumour immunity with therapeutic effect.
        Immunology. 2014; 143: 428-437
        • Hernández-Luna M.A.
        • Luria-Pérez R.
        Cancer immunotherapy: priming the host immune response with live attenuated Salmonella enterica.
        J Immunol Res. 2018; 2018: 2984247
        • Hervieu A.
        • Rébé C.
        • Végran F.
        • Chalmin F.
        • Bruchard M.
        • Vabres P.
        • et al.
        Dacarbazine-mediated upregulation of NKG2D ligands on tumor cells activates NK and CD8 T cells and restrains melanoma growth.
        J Invest Dermatol. 2013; 133: 499-508
        • Hodi F.S.
        • O'Day S.J.
        • McDermott D.F.
        • Weber R.W.
        • Sosman J.A.
        • Haanen J.B.
        • et al.
        Improved survival with ipilimumab in patients with metastatic melanoma [published correction appears in N Engl J Med 2010;363:1290].
        N Engl J Med. 2010; 363: 711-723
        • Hoiseth S.K.
        • Stocker B.A.
        Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines.
        Nature. 1981; 291: 238-239
        • Jenkins R.W.
        • Fisher D.E.
        Treatment of advanced melanoma in 2020 and beyond.
        J Invest Dermatol. 2021; 141: 23-31
        • Jin J.L.
        • Gong J.
        • Yin T.J.
        • Lu Y.J.
        • Xia J.J.
        • Xie Y.Y.
        • et al.
        PTD4-apoptin protein and dacarbazine show a synergistic antitumor effect on B16-F1 melanoma in vitro and in vivo.
        Eur J Pharmacol. 2011; 654: 17-25
        • Kramer M.G.
        • Masner M.
        • Casales E.
        • Moreno M.
        • Smerdou C.
        • Chabalgoity J.A.
        Neoadjuvant administration of Semliki Forest virus expressing interleukin-12 combined with attenuated Salmonella eradicates breast cancer metastasis and achieves long-term survival in immunocompetent mice.
        BMC Cancer. 2015; 15: 620
        • Najar H.M.
        • Dutz J.P.
        Topical CpG enhances the response of murine malignant melanoma to dacarbazine.
        J Invest Dermatol. 2008; 128: 2204-2210
        • Robert C.
        • Ribas A.
        • Wolchok J.D.
        • Hodi F.S.
        • Hamid O.
        • Kefford R.
        • et al.
        Anti-programmed-death-receptor-1 treatment with pembrolizumab in ipilimumab-refractory advanced melanoma: a randomised dose-comparison cohort of a phase 1 trial.
        Lancet. 2014; 384: 1109-1117
        • Serrone L.
        • Zeuli M.
        • Sega F.M.
        • Cognetti F.
        Dacarbazine-based chemotherapy for metastatic melanoma: thirty-year experience overview.
        J Exp Clin Cancer Res. 2000; 19: 21-34
        • Shah D.J.
        • Dronca R.S.
        Latest advances in chemotherapeutic, targeted, and immune approaches in the treatment of metastatic melanoma.
        Mayo Clin Proc. 2014; 89: 504-519
        • Sun J.
        • Kirichenko D.A.
        • Zager J.S.
        • Eroglu Z.
        The emergence of neoadjuvant therapy in advanced melanoma.
        Melanoma Manag. 2019; 6: MMT27
        • Tan K.W.
        • Evrard M.
        • Tham M.
        • Hong M.
        • Huang C.
        • Kato M.
        • et al.
        Tumor stroma and chemokines control T-cell migration into melanoma following temozolomide treatment.
        Oncoimmunology. 2015; 4e978709
        • Toso J.F.
        • Gill V.J.
        • Hwu P.
        • Marincola F.M.
        • Restifo N.P.
        • Schwartzentruber D.J.
        • et al.
        Phase I study of the intravenous administration of attenuated Salmonella typhimurium to patients with metastatic melanoma.
        J Clin Oncol. 2002; 20: 142-152
        • Ugurel S.
        • Paschen A.
        • Becker J.C.
        Dacarbazine in melanoma: from a chemotherapeutic drug to an immunomodulating agent.
        J Invest Dermatol. 2013; 133: 289-292
        • Vola M.
        • Mónaco A.
        • Bascuas T.
        • Rimsky G.
        • Agorio C.I.
        • Chabalgoity J.A.
        • et al.
        TLR7 agonist in combination with Salmonella as an effective antimelanoma immunotherapy.
        Immunotherapy. 2018; 10: 665-679
        • Wilson M.A.
        • Schuchter L.M.
        Chemotherapy for melanoma.
        Cancer Treat Res. 2016; 167: 209-229
        • Wolf M.
        • Eskerski H.
        • Bauder-Wüst U.
        • Haberkorn U.
        • Eisenhut M.
        Alkylating benzamides with melanoma cytotoxicity: experimental chemotherapy in a mouse melanoma model.
        Melanoma Res. 2006; 16: 487-496
        • Yang M.
        • Xu J.
        • Wang Q.
        • Zhang A.Q.
        • Wang K.
        An obligatory anaerobic Salmonella typhimurium strain redirects M2 macrophages to the M1 phenotype.
        Oncol Lett. 2018; 15: 3918-3922
        • Zhou S.
        • Gravekamp C.
        • Bermudes D.
        • Liu K.
        Tumour-targeting bacteria engineered to fight cancer.
        Nat Rev Cancer. 2018; 18: 727-743