Optimized biodegradable polymeric reservoir‐mediated local and sustained co‐delivery of dendritic cells and oncolytic adenovirus co‐expressing IL‐12 and GM‐CSF for cancer immunotherapy

ABSTRACT Administration of dendritic cells (DCs) combined with oncolytic adenovirus (Ad) expressing antitumor cytokines induces a potent antitumor effect and antitumor immunity by ameliorating the immunosuppressive tumor microenvironment. However, this combination therapy has significant limitations due to rapid dissemination and inactivation of the therapeutics at the tumor site, necessitating multiple injections of both therapeutics. To overcome these limitations, we have utilized gelatin‐based hydrogel to co‐deliver oncolytic Ad co‐expressing interleukin (IL)‐12 and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) (oAd) and DCs for sustained release of both therapeutics. The injectable and biodegradable hydrogels were prepared by mixing the polymer solutions containing horseradish peroxidase and hydrogen peroxide. Gel matrix enabled sustained release of both oAd and DCs while preserving their biological activity over a considerable time period, leading to efficient retention of both therapeutics in tumor tissue. Further, tumors treated with oAd‐ and DC‐loaded gel (oAd + DC/gel) showed a significantly greater expression level of IL‐12, GM‐CSF, and interferon‐&ggr; (IFN‐&ggr;) than either single treatment (oAd or DC) or oAd in combination with DC (oAd + DC), resulting in efficient activation of both endogenous and exogenous DCs, migration of DCs to draining lymph nodes, and tumor infiltration of CD4+ and CD8+ T cells. Moreover, oAd + DC/gel resulted in a significantly higher number of tumor‐specific IFN‐&ggr;–secreting immune cells compared with oAd + DC. Lastly, oAd + DC/gel significantly attenuated tumor‐mediated thymic atrophy, which is associated with immunosuppression in the tumor microenvironment, compared with oAd + DC. Taken together, these results demonstrate that gelatin gel‐mediated co‐delivery of oncolytic Ad and DCs might be a promising strategy to efficiently retain both therapeutics in tumor tissue and induce a potent antitumor immune response for an extended time period via a single administration.

[1]  R. Carrio,et al.  Impaired thymopoiesis occurring during the thymic involution of tumor-bearing mice is associated with a down-regulation of the antiapoptotic proteins Bcl-XL and A1. , 2009, International journal of molecular medicine.

[2]  H. Sung,et al.  In Situ Crosslinkable Gelatin Hydrogels for Vasculogenic Induction and Delivery of Mesenchymal Stem Cells , 2014, Advanced functional materials.

[3]  J. Patard,et al.  Dendritic cells are defective in breast cancer patients: a potential role for polyamine in this immunodeficiency , 2005, Breast Cancer Research.

[4]  Y. Bae,et al.  DC immunotherapy is highly effective for the inhibition of tumor metastasis or recurrence, although it is not efficient for the eradication of established solid tumors , 2007, Cancer Immunology, Immunotherapy.

[5]  Z. Berneman,et al.  Clinical use of dendritic cells for cancer therapy. , 2014, The Lancet. Oncology.

[6]  K. Park,et al.  Enzyme-catalyzed in situ forming gelatin hydrogels as bioactive wound dressings: effects of fibroblast delivery on wound healing efficacy. , 2014, Journal of materials chemistry. B.

[7]  Yu Seong Lee,et al.  Concurrent delivery of GM-CSF and B7-1 using an oncolytic adenovirus elicits potent antitumor effect , 2006, Gene Therapy.

[8]  B. Han,et al.  Optimizing dendritic cell-based immunotherapy for cancer , 2007, Expert review of vaccines.

[9]  B. Fang,et al.  Oncolytic virotherapy for cancer treatment: challenges and solutions , 2005, The journal of gene medicine.

[10]  V. Cerundolo,et al.  Dendritic cells: a journey from laboratory to clinic , 2004, Nature Immunology.

[11]  E. Marshall.  Cancer Trial of Interleukin-12 Halted , 1995 .

[12]  J. Leonard,et al.  Effects of Single-Dose Interleukin-12 Exposure on Interleukin-12–Associated Toxicity and Interferon-γ Production , 1997 .

[13]  C. Kim,et al.  Antitumor effects of combined granulocyte macrophage colony stimulating factor and macrophage inflammatory protein‐3 alpha plasmid DNA , 2010, Cancer science.

[14]  G. Dranoff,et al.  Cytokines in cancer pathogenesis and cancer therapy , 2004, Nature Reviews Cancer.

[15]  C. Yun,et al.  Strengthening of antitumor immune memory and prevention of thymic atrophy mediated by adenovirus expressing IL-12 and GM-CSF , 2011, Gene Therapy.

[16]  D. Irvine,et al.  Injectable dendritic cell-carrying alginate gels for immunization and immunotherapy. , 2008, Biomaterials.

[17]  G. Fuller,et al.  Expression and localization of 72 kDa type IV collagenase (MMP-2) in human malignant gliomas in vivo , 2004, Clinical and Experimental Metastasis.

[18]  R. Crystal,et al.  Innate immune mechanisms dominate elimination of adenoviral vectors following in vivo administration. , 1997, Human gene therapy.

[19]  C. Yun,et al.  Oncolytic adenovirus co-expressing IL-12 and IL-18 improves tumor-specific immunity via differentiation of T cells expressing IL-12Rβ2 or IL-18Rα , 2011, Gene Therapy.

[20]  C. Yun,et al.  Oncolytic Adenovirus Expressing IL-23 and p35 Elicits IFN-γ- and TNF-α-Co-Producing T Cell-Mediated Antitumor Immunity , 2013, PloS one.

[21]  Wei Zhu,et al.  Immune response, clinical outcome and safety of dendritic cell vaccine in combination with cytokine-induced killer cell therapy in cancer patients , 2013, Oncology letters.

[22]  J. Prieto,et al.  Gene therapy of cancer based on interleukin 12. , 2005, Current gene therapy.

[23]  A. Palucka,et al.  Dendritic cells as therapeutic vaccines against cancer , 2005, Nature Reviews Immunology.

[24]  M. Woodruff,et al.  Immune Response , 1969, Nature.

[25]  T. Kubota,et al.  A phase I/II clinical trial investigating the adverse and therapeutic effects of a postoperative autologous dendritic cell tumor vaccine in patients with malignant glioma , 2011, Journal of Clinical Neuroscience.

[26]  Hoguen Kim,et al.  Therapeutic and Tumor-specific Immunity Induced by Combination of Dendritic Cells and Oncolytic Adenovirus Expressing IL-12 and 4-1BBL. , 2010, Molecular therapy : the journal of the American Society of Gene Therapy.

[27]  C. Yun,et al.  Optimizing DC Vaccination by Combination With Oncolytic Adenovirus Coexpressing IL-12 and GM-CSF , 2011, Molecular therapy : the journal of the American Society of Gene Therapy.

[28]  C. Figdor,et al.  Dendritic Cell–Based Immunotherapy: State of the Art and Beyond , 2016, Clinical Cancer Research.

[29]  Peter Brossart,et al.  Immunologic and clinical responses after vaccinations with peptide-pulsed dendritic cells in metastatic renal cancer patients. , 2006, Cancer research.

[30]  Hoguen Kim,et al.  Relaxin expression from tumor-targeting adenoviruses and its intratumoral spread, apoptosis induction, and efficacy. , 2006, Journal of the National Cancer Institute.

[31]  G. Dranoff,et al.  Gene therapy for metastatic brain tumors by vaccination with granulocyte-macrophage colony-stimulating factor-transduced tumor cells. , 1997, Human gene therapy.

[32]  G. William,et al.  Cellular Mechanism of Thymic Involution , 2003, Scandinavian journal of immunology.

[33]  Hoguen Kim,et al.  Enhanced Antitumor Effect of Oncolytic Adenovirus Expressing Interleukin-12 and B7-1 in an Immunocompetent Murine Model , 2006, Clinical Cancer Research.

[34]  C Caux,et al.  The cytokine profile expressed by human dendritic cells is dependent on cell subtype and mode of activation. , 1998, Journal of immunology.

[35]  H. Ragde,et al.  Follow‐up evaluation of prostate cancer patients infused with autologous dendritic cells pulsed with PSMA peptides , 1997, The Prostate.

[36]  E. Kang,et al.  Local sustained delivery of oncolytic adenovirus with injectable alginate gel for cancer virotherapy , 2013, Gene Therapy.

[37]  H. Okamura,et al.  A mechanism underlying synergy between IL-12 and IFN-gamma-inducing factor in enhanced production of IFN-gamma. , 1997, Journal of immunology.

[38]  K. Schäkel,et al.  Reciprocal activating interaction between 6‐sulfo LacNAc+ dendritic cells and NK cells , 2009, International journal of cancer.

[39]  J. Leonard,et al.  Effects of single-dose interleukin-12 exposure on interleukin-12-associated toxicity and interferon-gamma production. , 1997, Blood.

[40]  R. Ramesh,et al.  Cytokine- and chemokine-based gene therapy for cancer. , 2003, Current opinion in molecular therapeutics.

[41]  G. Wolff,et al.  Role of alveolar macrophages in rapid elimination of adenovirus vectors administered to the epithelial surface of the respiratory tract. , 1997, Human gene therapy.

[42]  O. Podhajcer,et al.  Cytokine gene transfer for cancer therapy. , 2007, Cytokine & growth factor reviews.

[43]  Dirk Schadendorf,et al.  Vaccination of melanoma patients with peptide- or tumorlysate-pulsed dendritic cells , 1998, Nature Medicine.

[44]  Edgar G. Engleman,et al.  Vaccination of patients with B–cell lymphoma using autologous antigen–pulsed dendritic cells , 1996, Nature Medicine.

[45]  S. Szabo,et al.  Molecular mechanisms regulating Th1 immune responses. , 2003, Annual review of immunology.

[46]  Y. Yonemitsu,et al.  Clinical and Immunologic Evaluation of Dendritic Cell–Based Immunotherapy in Combination With Gemcitabine and/or S-1 in Patients With Advanced Pancreatic Carcinoma , 2012, Pancreas.