Enhanced targeting of triple-negative breast carcinoma and malignant melanoma by photochemical internalization of CSPG4-targeting immunotoxins

Triple-negative breast cancer (TNBC) and malignant melanoma are highly aggressive cancers that widely express the cell surface chondroitin sulfate proteoglycan 4 (CSPG4/NG2). CSPG4 plays an important role in tumor cell growth and survival and promotes chemo- and radiotherapy resistance, suggesting that CSPG4 is an attractive target in cancer therapy. In the present work, we applied the drug delivery technology photochemical internalization (PCI) in combination with the novel CSPG4-targeting immunotoxin 225.28-saporin as an efficient and specific strategy to kill aggressive TNBC and amelanotic melanoma cells. Light-activation of the clinically relevant photosensitizer TPCS2a (fimaporfin) and 225.28-saporin was found to act in a synergistic manner, and was superior to both PCI of saporin and PCI-no-drug (TPCS2a + light only) in three TNBC cell lines (MDA-MB-231, MDA-MB-435 and SUM149) and two BRAFV600E mutated malignant melanoma cell lines (Melmet 1 and Melmet 5). The cytotoxic effect was highly dependent on the light dose and expression of CSPG4 since no enhanced cytotoxicity of PCI of 225.28-saporin compared to PCI of saporin was observed in the CSPG4-negative MCF-7 cells. The PCI of a smaller, and clinically relevant CSPG4-targeting toxin (scFvMEL-rGel) validated the CSPG4-targeting concept in vitro and induced a strong inhibition of tumor growth in the amelanotic melanoma xenograft A-375 model. In conclusion, the combination of the drug delivery technology PCI and CSPG4-targeting immunotoxins is an efficient, specific and light-controlled strategy for the elimination of aggressive cells of TNBC and malignant melanoma origin. This study lays the foundation for further preclinical evaluation of PCI in combination with CSPG4-targeting.

[1]  G. Brede,et al.  Photochemical Internalization of Peptide Antigens Provides a Novel Strategy to Realize Therapeutic Cancer Vaccination , 2018, Front. Immunol..

[2]  K. Berg,et al.  Development of resistance to photodynamic therapy (PDT) in human breast cancer cells is photosensitizer‐dependent: Possible mechanisms and approaches for overcoming PDT‐resistance , 2017, Biochemical pharmacology.

[3]  K. Berg,et al.  Disulfonated tetraphenyl chlorin (TPCS2a)-induced photochemical internalisation of bleomycin in patients with solid malignancies: a phase 1, dose-escalation, first-in-man trial. , 2016, The Lancet. Oncology.

[4]  V. LeBleu,et al.  EMT Program is Dispensable for Metastasis but Induces Chemoresistance in Pancreatic Cancer , 2015, Nature.

[5]  K. Berg,et al.  Photochemical activation of drugs for the treatment of therapy-resistant cancers , 2015, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[6]  K. Berg,et al.  Photochemical internalisation, a minimally invasive strategy for light-controlled endosomal escape of cancer stem cell-targeting therapeutics , 2015, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[7]  Q. Peng,et al.  Light-controlled endosomal escape of the novel CD133-targeting immunotoxin AC133-saporin by photochemical internalization - A minimally invasive cancer stem cell-targeting strategy. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[8]  T. Kündig,et al.  Photosensitisation facilitates cross-priming of adjuvant-free protein vaccines and stimulation of tumour-suppressing CD8 T cells. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[9]  T. Graeber,et al.  Low MITF/AXL ratio predicts early resistance to multiple targeted drugs in melanoma , 2014, Nature Communications.

[10]  K. Berg,et al.  Light-triggered, efficient cytosolic release of IM7-saporin targeting the putative cancer stem cell marker CD44 by photochemical internalization. , 2014, Molecular pharmaceutics.

[11]  J. Mesirov,et al.  A melanoma cell state distinction influences sensitivity to MAPK pathway inhibitors. , 2014, Cancer discovery.

[12]  S. Krauss,et al.  The novel EpCAM-targeting monoclonal antibody 3–17I linked to saporin is highly cytotoxic after photochemical internalization in breast, pancreas and colon cancer cell lines , 2014, mAbs.

[13]  T. Kündig,et al.  Intradermal photosensitisation facilitates stimulation of MHC class-I restricted CD8 T-cell responses of co-administered antigen. , 2014, Journal of controlled release : official journal of the Controlled Release Society.

[14]  E. Hovig,et al.  Melanoma brain colonization involves the emergence of a brain-adaptive phenotype , 2014, Oncoscience.

[15]  L. Polito,et al.  Saporin-S6: A Useful Tool in Cancer Therapy , 2013, Toxins.

[16]  K. Berg,et al.  Photochemical internalization of CD133-targeting immunotoxins efficiently depletes sarcoma cells with stem-like properties and reduces tumorigenicity. , 2013, Biochimica et biophysica acta.

[17]  W. Yang,et al.  Spatiotemporally controlled induction of autophagy-mediated lysosome turnover , 2013, Nature Communications.

[18]  K. Berg,et al.  Photochemical internalization (PCI) of immunotoxins targeting CD133 is specific and highly potent at femtomolar levels in cells with cancer stem cell properties. , 2013, Journal of controlled release : official journal of the Controlled Release Society.

[19]  Michael R Hamblin,et al.  Melanoma resistance to photodynamic therapy: new insights , 2013, Biological chemistry.

[20]  J. Panyam,et al.  Immunotoxin targeting CD133+ breast carcinoma cells , 2012, Drug Delivery and Translational Research.

[21]  Kristian Berg,et al.  Photochemical internalization (PCI) of HER2-targeted toxins: synergy is dependent on the treatment sequence. , 2012, Biochimica et biophysica acta.

[22]  K. Flaherty,et al.  From genes to drugs: targeted strategies for melanoma , 2012, Nature Reviews Cancer.

[23]  K. Berg,et al.  Strongly amphiphilic photosensitizers are not substrates of the cancer stem cell marker ABCG2 and provides specific and efficient light-triggered drug delivery of an EGFR-targeted cytotoxic drug. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[24]  H. Pass,et al.  CSPG4 as a Target of Antibody-Based Immunotherapy for Malignant Mesothelioma , 2012, Clinical Cancer Research.

[25]  Alysha K Croker,et al.  Inhibition of aldehyde dehydrogenase (ALDH) activity reduces chemotherapy and radiation resistance of stem-like ALDHhiCD44+ human breast cancer cells , 2012, Breast Cancer Research and Treatment.

[26]  Simon C Watkins,et al.  Functional characterization of an scFv-Fc antibody that immunotherapeutically targets the common cancer cell surface proteoglycan CSPG4. , 2011, Cancer research.

[27]  Gui-yuan Li,et al.  CSPG4, a potential therapeutic target, facilitates malignant progression of melanoma , 2011, Pigment cell & melanoma research.

[28]  Paul Ellis,et al.  Molecular heterogeneity of triple-negative breast cancer and its clinical implications , 2011, Current opinion in oncology.

[29]  Even Angell-Petersen,et al.  Disulfonated tetraphenyl chlorin (TPCS_2a), a novel photosensitizer developed for clinical utilization of photochemical internalization , 2011, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[30]  Kristian Berg,et al.  Photochemical internalization of tumor‐targeted protein toxins , 2011, Lasers in surgery and medicine.

[31]  H. Immervoll,et al.  Expression of the progenitor marker NG2/CSPG4 predicts poor survival and resistance to ionising radiation in glioblastoma , 2011, Acta Neuropathologica.

[32]  K. Reddy,et al.  Triple-negative breast cancers: an updated review on treatment options. , 2011, Current oncology.

[33]  H. Immervoll,et al.  Targeting the NG2/CSPG4 Proteoglycan Retards Tumour Growth and Angiogenesis in Preclinical Models of GBM and Melanoma , 2011, PloS one.

[34]  X. Chen,et al.  Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. , 2011, The Journal of clinical investigation.

[35]  Axel Hoos,et al.  Ipilimumab plus dacarbazine for previously untreated metastatic melanoma. , 2011, The New England journal of medicine.

[36]  R. Dummer,et al.  A proliferative melanoma cell phenotype is responsive to RAF/MEK inhibition independent of BRAF mutation status , 2011, Pigment cell & melanoma research.

[37]  S. Nelson,et al.  Melanomas acquire resistance to B-RAF(V600E) inhibition by RTK or N-RAS upregulation , 2010, Nature.

[38]  K. Hoek,et al.  Cancer stem cells versus phenotype‐switching in melanoma , 2010, Pigment cell & melanoma research.

[39]  K. Berg,et al.  Photochemical internalization provides time- and space-controlled endolysosomal escape of therapeutic molecules. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[40]  W. Barry,et al.  CSPG4 protein as a new target for the antibody-based immunotherapy of triple-negative breast cancer. , 2010, Journal of the National Cancer Institute.

[41]  Jason I. Herschkowitz,et al.  Phenotypic and molecular characterization of the claudin-low intrinsic subtype of breast cancer , 2010, Breast Cancer Research.

[42]  D. Schadendorf,et al.  Improved survival with ipilimumab in patients with metastatic melanoma. , 2010, The New England journal of medicine.

[43]  W. Woodward,et al.  Overcoming radiation resistance in inflammatory breast cancer , 2010, Cancer.

[44]  S. Aamdal,et al.  Human malignant melanoma harbours a large fraction of highly clonogenic cells that do not express markers associated with cancer stem cells , 2010, Pigment cell & melanoma research.

[45]  S. Ferrone,et al.  Functional and clinical relevance of chondroitin sulfate proteoglycan 4. , 2010, Advances in cancer research.

[46]  King-Jen Chang,et al.  Multiple Lineages of Human Breast Cancer Stem/Progenitor Cells Identified by Profiling with Stem Cell Markers , 2009, PloS one.

[47]  N Harbeck,et al.  Triple-negative breast cancer--current status and future directions. , 2009, Annals of oncology : official journal of the European Society for Medical Oncology.

[48]  K. Berg,et al.  Multi-Modality Therapeutics with Potent Anti-Tumor Effects: Photochemical Internalization Enhances Delivery of the Fusion Toxin scFvMEL/rGel , 2009, PloS one.

[49]  A. Chambers,et al.  MDA-MB-435 and M14 cell lines: identical but not M14 melanoma? , 2009, Cancer research.

[50]  Charlotte Kuperwasser,et al.  Human breast cancer cell lines contain stem-like cells that self-renew, give rise to phenotypically diverse progeny and survive chemotherapy , 2008, Breast Cancer Research.

[51]  S. Narod,et al.  Triple-Negative Breast Cancer: Clinical Features and Patterns of Recurrence , 2007, Clinical Cancer Research.

[52]  R. Marais,et al.  Melanoma biology and new targeted therapy , 2007, Nature.

[53]  K. Berg,et al.  Targeted delivery and enhanced cytotoxicity of cetuximab-saporin by photochemical internalization in EGFR-positive cancer cells. , 2007, Molecular pharmaceutics.

[54]  Chad J. Creighton,et al.  MDA-MB-435 cells are derived from M14 Melanoma cells––a loss for breast cancer, but a boon for melanoma research , 2007, Breast Cancer Research and Treatment.

[55]  Frank Pajonk,et al.  The response of CD24(-/low)/CD44+ breast cancer-initiating cells to radiation. , 2006, Journal of the National Cancer Institute.

[56]  K. Berg,et al.  Reversal of doxorubicin resistance in breast cancer cells by photochemical internalization , 2006, International journal of cancer.

[57]  K. Berg,et al.  Photochemical Internalization of Therapeutic Macromolecular Agents: A Novel Strategy to Kill Multidrug-Resistant Cancer Cells , 2006, Journal of Pharmacology and Experimental Therapeutics.

[58]  Michael R Hamblin,et al.  Photodynamic therapy and anti-tumour immunity , 2006, Nature Reviews Cancer.

[59]  K. Berg,et al.  Photochemically stimulated drug delivery increases the cytotoxicity and specificity of EGF-saporin. , 2006, Journal of Controlled Release.

[60]  J. Haveman,et al.  Clonogenic assay of cells in vitro , 2006, Nature Protocols.

[61]  M. Weichenthal,et al.  Fotemustine compared with dacarbazine in patients with disseminated malignant melanoma: a phase III study. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[62]  S. Ferrone,et al.  Human high molecular weight-melanoma-associated antigen (HMW-MAA): a melanoma cell surface chondroitin sulfate proteoglycan (MSCP) with biological and clinical significance. , 2004, Critical reviews in immunology.

[63]  F. Watt,et al.  Role of melanoma chondroitin sulphate proteoglycan in patterning stem cells in human interfollicular epidermis , 2003, Development.

[64]  M. Rosenblum,et al.  Design, expression, purification, and characterization, in vitro and in vivo, of an antimelanoma single-chain Fv antibody fused to the toxin gelonin. , 2003, Cancer research.

[65]  S. Morrison,et al.  Prospective identification of tumorigenic breast cancer cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[66]  K. Berg,et al.  5-Aminolevulinic Acid–based Photochemical Internalization of the Immunotoxin MOC31-gelonin Generates Synergistic Cytotoxic Effects In Vitro¶ , 2001, Photochemistry and photobiology.

[67]  K. Berg,et al.  Photochemical internalisation increases the cytotoxic effect of the immunotoxin MOC31‐gelonin , 2000, International journal of cancer.

[68]  S. Merajver,et al.  A novel putative low-affinity insulin-like growth factor-binding protein, LIBC (lost in inflammatory breast cancer), and RhoC GTPase correlate with the inflammatory breast cancer phenotype. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[69]  H Anholt,et al.  Photochemical internalization: a novel technology for delivery of macromolecules into cytosol. , 1999, Cancer research.

[70]  D. Kessel,et al.  TUMOR LOCALIZATION AND PHOTOSENSITIZATION BY SULFONATED DERIVATIVES OF TETRAPHENYLPORPHINE , 1987, Photochemistry and photobiology.

[71]  S. Ferrone,et al.  Distribution and molecular characterization of a cell‐surface and a cytoplasmic antigen detectable in human melanoma cells with monoclonal antibodies , 1981, International journal of cancer.

[72]  G. Steel,et al.  Exploitable mechanisms in combined radiotherapy-chemotherapy: the concept of additivity. , 1979, International journal of radiation oncology, biology, physics.