Journal Article
. 2021 Jul; 13(13):.
doi: 10.3390/cancers13133356.

Evaluation of In Vitro Phototoxicity of a Minibody-IR700 Conjugate Using Cell Monolayer and Multicellular Tumor Spheroid Models

Mouldy Sioud 1 Petras Juzenas 2 Qindong Zhang 1 Andrius Kleinauskas 2 Qian Peng 2 
  • PMID: 34283089
  •     50 References
  •     1 citations


Photodynamic therapy (PDT) is a treatment strategy that utilizes photosensitizers (PSs) and light of a specific wavelength to kill cancer cells. However, limited tumor specificity is still a drawback for the clinical application of PDT. To increase the therapeutic efficacy and specificity of PDT, a novel human minibody (MS5) that recognizes a cell surface receptor expressed on various cancer cells was labeled with the hydrophilic phthalocyanine PS IR700 to generate an MS5-IR700 conjugate that is activated by near-infrared (NIR) light. The phototoxicity of the conjugate was mainly tested against the PC3 prostate cancer cell line. The MS5-IR700 conjugate killed PC3 cells after NIR light irradiation as compared to untreated cells or cells treated with IR700 alone. Time-course analysis of cell viability revealed a high percentage of cell death during the first hour in PC3 cells exposed to the MS5-IR700 conjugate and NIR light irradiation. After irradiation, the MS5-IR700 conjugate-treated PC3 cells displayed cellular swelling, round shape, and rupture of the cell and nuclear membranes. In a co-culture model, the MS5-IR700 conjugate killed MS5-positive Ramos lymphoma cells specifically, while leaving MS5-negative cells unaffected. In line with the data obtained with the monolayer cultures, the MS5-IR700 conjugate also killed PC3 cancer cell spheroids. The treatment induced relocation of heat shock protein 70 and calreticulin to the cell surface, implying the induction of immunogenic cell death. Overall, the data suggest that the developed MS5-IR700 conjugate is a promising therapeutic agent that warrants further preclinical studies.

Keywords: antibodies; antibody-drug conjugates; photodynamic therapy; photoimmunotherapy; photosensitizers.

Photodynamic therapy.
T J Dougherty, C J Gomer, +5 authors, Q Peng.
J Natl Cancer Inst, 1998 Jun 24; 90(12). PMID: 9637138    Free PMC article.
Highly Cited. Review.
Oncologic Photodynamic Therapy: Basic Principles, Current Clinical Status and Future Directions.
Demian van Straten, Vida Mashayekhi, +2 authors, Dominic J Robinson.
Cancers (Basel), 2017 Feb 22; 9(2). PMID: 28218708    Free PMC article.
Highly Cited. Review.
PDGFRβ-specific affibody-directed delivery of a photosensitizer, IR700, is efficient for vascular-targeted photodynamic therapy of colorectal cancer.
Qiuxiao Shi, Ze Tao, +4 authors, Xiaofeng Lu.
Drug Deliv, 2017 Nov 29; 24(1). PMID: 29182023    Free PMC article.
Heat-shock proteins in diagnosis and treatment: an overview of different biochemical and immunological functions.
Alireza Milani, Marzieh Basirnejad, Azam Bolhassani.
Immunotherapy, 2019 Feb 08; 11(3). PMID: 30730280
A novel pathway combining calreticulin exposure and ATP secretion in immunogenic cancer cell death.
Abhishek D Garg, Dmitri V Krysko, +12 authors, Patrizia Agostinis.
EMBO J, 2012 Jan 19; 31(5). PMID: 22252128    Free PMC article.
Highly Cited.
Top 10 Challenges in Cancer Immunotherapy.
Priti S Hegde, Daniel S Chen.
Immunity, 2020 Jan 16; 52(1). PMID: 31940268
Immunogenic cell death in cancer therapy.
Guido Kroemer, Lorenzo Galluzzi, Oliver Kepp, Laurence Zitvogel.
Annu Rev Immunol, 2012 Nov 20; 31. PMID: 23157435
Highly Cited. Review.
Recent advances in strategies for overcoming hypoxia in photodynamic therapy of cancer.
Barbara Pucelik, Adam Sułek, Agata Barzowska, Janusz M Dąbrowski.
Cancer Lett, 2020 Jul 22; 492. PMID: 32693200
Spherical cancer models in tumor biology.
Louis-Bastien Weiswald, Dominique Bellet, Virginie Dangles-Marie.
Neoplasia, 2015 Jan 28; 17(1). PMID: 25622895    Free PMC article.
Highly Cited. Review.
Phage Display Libraries: From Binders to Targeted Drug Delivery and Human Therapeutics.
Mouldy Sioud.
Mol Biotechnol, 2019 Feb 08; 61(4). PMID: 30729435
Early and Late Onset Side Effects of Photodynamic Therapy.
Francesco Borgia, Roberta Giuffrida, +3 authors, Serafinella P Cannavò.
Biomedicines, 2018 Feb 01; 6(1). PMID: 29382133    Free PMC article.
Intratumoral depletion of regulatory T cells using CD25-targeted photodynamic therapy in a mouse melanoma model induces antitumoral immune responses.
Dong Sun Oh, Heegon Kim, +4 authors, Heung Kyu Lee.
Oncotarget, 2017 May 26; 8(29). PMID: 28537894    Free PMC article.
Major apoptotic mechanisms and genes involved in apoptosis.
Yağmur Kiraz, Aysun Adan, Melis Kartal Yandim, Yusuf Baran.
Tumour Biol, 2016 Apr 10; 37(7). PMID: 27059734
Highly Cited. Review.
Cell Death Mechanisms in Tumoral and Non-Tumoral Human Cell Lines Triggered by Photodynamic Treatments: Apoptosis, Necrosis and Parthanatos.
J Soriano, I Mora-Espí, +4 authors, C Nogués.
Sci Rep, 2017 Jan 24; 7. PMID: 28112275    Free PMC article.
Syngeneic Mouse Models of Oral Cancer Are Effectively Targeted by Anti-CD44-Based NIR-PIT.
Tadanobu Nagaya, Yuko Nakamura, +5 authors, Hisataka Kobayashi.
Mol Cancer Res, 2017 Sep 20; 15(12). PMID: 28923838    Free PMC article.
Stem cells in cancer therapy: opportunities and challenges.
Cheng-Liang Zhang, Ting Huang, +2 authors, Dong Liu.
Oncotarget, 2017 Nov 02; 8(43). PMID: 29088907    Free PMC article.
Integrin Signaling in Cancer: Mechanotransduction, Stemness, Epithelial Plasticity, and Therapeutic Resistance.
Jonathan Cooper, Filippo G Giancotti.
Cancer Cell, 2019 Mar 20; 35(3). PMID: 30889378    Free PMC article.
Highly Cited. Review.
An anti-leukemic single chain Fv antibody selected from a synthetic human phage antibody library.
M Shadidi, M Sioud.
Biochem Biophys Res Commun, 2001 Feb 13; 280(2). PMID: 11162553
Photodynamic therapy for cancer.
Dennis E J G J Dolmans, Dai Fukumura, Rakesh K Jain.
Nat Rev Cancer, 2003 May 02; 3(5). PMID: 12724736
Highly Cited. Review.
A Novel Theranostic Combination of Near-infrared Fluorescence Imaging and Laser Irradiation Targeting c-KIT for Gastrointestinal Stromal Tumors.
Shota Fujimoto, Naoki Muguruma, +8 authors, Tetsuji Takayama.
Theranostics, 2018 May 04; 8(9). PMID: 29721082    Free PMC article.
Vimentin in cancer and its potential as a molecular target for cancer therapy.
Arun Satelli, Shulin Li.
Cell Mol Life Sci, 2011 Jun 04; 68(18). PMID: 21637948    Free PMC article.
Highly Cited. Review.
Cancer immunoediting and resistance to T cell-based immunotherapy.
Jake S O'Donnell, Michele W L Teng, Mark J Smyth.
Nat Rev Clin Oncol, 2018 Dec 12; 16(3). PMID: 30523282
Highly Cited. Review.
Stimulation of dendritic cells by DAMPs in ALA-PDT treated SCC tumor cells.
Xiaojie Wang, Jie Ji, +7 authors, Xiuli Wang.
Oncotarget, 2015 Dec 02; 6(42). PMID: 26625309    Free PMC article.
The Potential of Nanobody-Targeted Photodynamic Therapy to Trigger Immune Responses.
Irati Beltrán Hernández, Mathieu L Angelier, +3 authors, Sabrina Oliveira.
Cancers (Basel), 2020 Apr 25; 12(4). PMID: 32326519    Free PMC article.
Immunomodulatory activity of IR700-labelled affibody targeting HER2.
Justyna Mączyńska, Chiara Da Pieve, +5 authors, Gabriela Kramer-Marek.
Cell Death Dis, 2020 Oct 22; 11(10). PMID: 33082328    Free PMC article.
Cancer cell-selective in vivo near infrared photoimmunotherapy targeting specific membrane molecules.
Makoto Mitsunaga, Mikako Ogawa, +3 authors, Hisataka Kobayashi.
Nat Med, 2011 Nov 08; 17(12). PMID: 22057348    Free PMC article.
Highly Cited.
Mechanisms of resistance to trastuzumab emtansine (T-DM1) in HER2-positive breast cancer.
Francis W Hunter, Hilary R Barker, +5 authors, Stephen M F Jamieson.
Br J Cancer, 2019 Dec 17; 122(5). PMID: 31839676    Free PMC article.
Preclinical and Clinical Evidence of Immune Responses Triggered in Oncologic Photodynamic Therapy: Clinical Recommendations.
Irati Beltrán Hernández, Yingxin Yu, +2 authors, Sabrina Oliveira.
J Clin Med, 2020 Jan 30; 9(2). PMID: 31991650    Free PMC article.
The inducible Hsp70 as a marker of tumor immunogenicity.
P R Clark, A Ménoret.
Cell Stress Chaperones, 2001 Oct 16; 6(2). PMID: 11599573    Free PMC article.
Photodynamic therapy - mechanisms, photosensitizers and combinations.
Stanisław Kwiatkowski, Bartosz Knap, +7 authors, Julita Kulbacka.
Biomed Pharmacother, 2018 Aug 19; 106. PMID: 30119176
Highly Cited. Review.
Loss of CD20 antigen expression after rituximab therapy of CD20 positive B cell lymphoma (diffuse large B cell extranodal marginal zone lymphoma combination): a case report and review of the literature.
Berna Bozkurt Duman, Berksoy Sahin, Melek Ergin, Birol Guvenc.
Med Oncol, 2011 Aug 02; 29(2). PMID: 21805377
The Course of Immune Stimulation by Photodynamic Therapy: Bridging Fundamentals of Photochemically Induced Immunogenic Cell Death to the Enrichment of T-Cell Repertoire.
Shubhankar Nath, Girgis Obaid, Tayyaba Hasan.
Photochem Photobiol, 2019 Oct 12; 95(6). PMID: 31602649    Free PMC article.
Leveraging the immune system during chemotherapy: moving calreticulin to the cell surface converts apoptotic death from "silent" to immunogenic.
Michel Obeid, Theocharis Panaretakis, +6 authors, Guido Kroemer.
Cancer Res, 2007 Sep 07; 67(17). PMID: 17804698
Building better monoclonal antibody-based therapeutics.
George J Weiner.
Nat Rev Cancer, 2015 May 23; 15(6). PMID: 25998715    Free PMC article.
Highly Cited. Review.
Antibodies armed with photosensitizers: from chemical synthesis to photobiological applications.
Patricia M R Pereira, Barbara Korsak, +3 authors, João P C Tomé.
Org Biomol Chem, 2015 Jan 23; 13(9). PMID: 25612113
Epidermal Growth Factor Receptor (EGFR)-targeted Photoimmunotherapy (PIT) for the Treatment of EGFR-expressing Bladder Cancer.
Reema Railkar, L Spencer Krane, +8 authors, Piyush K Agarwal.
Mol Cancer Ther, 2017 Jun 18; 16(10). PMID: 28619755    Free PMC article.
Host Immunity Following Near-Infrared Photoimmunotherapy Is Enhanced with PD-1 Checkpoint Blockade to Eradicate Established Antigenic Tumors.
Tadanobu Nagaya, Jay Friedman, +6 authors, Hisataka Kobayashi.
Cancer Immunol Res, 2019 Jan 27; 7(3). PMID: 30683733    Free PMC article.
Lamin A/C cleavage by caspase-6 activation is crucial for apoptotic induction by photodynamic therapy with hexaminolevulinate in human B-cell lymphoma cells.
Susan Shahzidi, Andreas Brech, +4 authors, Qian Peng.
Cancer Lett, 2013 Aug 07; 339(1). PMID: 23916608
Photoinduced Ligand Release from a Silicon Phthalocyanine Dye Conjugated with Monoclonal Antibodies: A Mechanism of Cancer Cell Cytotoxicity after Near-Infrared Photoimmunotherapy.
Kazuhide Sato, Kanta Ando, +12 authors, Hisataka Kobayashi.
ACS Cent Sci, 2018 Dec 18; 4(11). PMID: 30555909    Free PMC article.
Cytokines in immunogenic cell death: Applications for cancer immunotherapy.
Anne Showalter, Arati Limaye, +4 authors, Annette R Khaled.
Cytokine, 2017 Jun 27; 97. PMID: 28648866    Free PMC article.
Immunogenic cancer cell death selectively induced by near infrared photoimmunotherapy initiates host tumor immunity.
Mikako Ogawa, Yusuke Tomita, +13 authors, Hisataka Kobayashi.
Oncotarget, 2017 Jan 07; 8(6). PMID: 28060726    Free PMC article.
Highly Cited.
Imaging and Selective Elimination of Glioblastoma Stem Cells with Theranostic Near-Infrared-Labeled CD133-Specific Antibodies.
Hua Jing, Claudia Weidensteiner, +5 authors, Gabriele Niedermann.
Theranostics, 2016 May 11; 6(6). PMID: 27162556    Free PMC article.
Near-infrared photoimmunotherapy of pancreatic cancer using an indocyanine green-labeled anti-tissue factor antibody.
Winn Aung, Atsushi B Tsuji, +4 authors, Tatsuya Higashi.
World J Gastroenterol, 2019 Jan 10; 24(48). PMID: 30622378    Free PMC article.
Near-infrared photoimmunotherapy of cancer: a new approach that kills cancer cells and enhances anti-cancer host immunity.
Hisataka Kobayashi, Aki Furusawa, Adrian Rosenberg, Peter L Choyke.
Int Immunol, 2020 Jun 05; 33(1). PMID: 32496557    Free PMC article.
Near infrared photoimmunotherapy targeting DLL3 for small cell lung cancer.
Yoshitaka Isobe, Kazuhide Sato, +14 authors, Yoshinori Hasegawa.
EBioMedicine, 2020 Jan 26; 52. PMID: 31981983    Free PMC article.
Epidermal growth factor receptor (EGFR) density may not be the only determinant for the efficacy of EGFR-targeted photoimmunotherapy in human head and neck cancer cell lines.
Wei Peng, Henriette S de Bruijn, +7 authors, Dominic J Robinson.
Lasers Surg Med, 2018 May 20; 50(5). PMID: 29777587
Photoimmunotherapy of Ovarian Cancer: A Unique Niche in the Management of Advanced Disease.
Shubhankar Nath, Mohammad Ahsan Saad, +2 authors, Tayyaba Hasan.
Cancers (Basel), 2019 Dec 01; 11(12). PMID: 31783651    Free PMC article.
Development of a new high-affinity human antibody with antitumor activity against solid and blood malignancies.
Mouldy Sioud, Phuong Westby, +2 authors, Qian Peng.
FASEB J, 2018 Jun 20; 32(9). PMID: 29913558
Photodynamic therapy (PDT) of cancer: from local to systemic treatment.
Janusz M Dąbrowski, Luis G Arnaut.
Photochem Photobiol Sci, 2015 Jul 30; 14(10). PMID: 26219737
EGFR Family Members' Regulation of Autophagy Is at a Crossroads of Cell Survival and Death in Cancer.
Elizabeth Henson, Yongqiang Chen, Spencer Gibson.
Cancers (Basel), 2017 Mar 25; 9(4). PMID: 28338617    Free PMC article.
Correction: Sioud et al. Evaluation of In Vitro Phototoxicity of a Minibody-IR700 Conjugate Using Monolayer and Multicellular Tumor Spheroid Models. Cancers 2021, 13, 3356.
Mouldy Sioud, Petras Juzenas, +2 authors, Qian Peng.
Cancers (Basel), 2021 Nov 14; 13(21). PMID: 34771754    Free PMC article.