A non-toxic approach for treatment of breast cancer and its metastases: capecitabine enhanced photodynamic therapy in a murine breast tumor model

Abstract

Aim: Breast cancer (BCA) in women is a leading cause of mortality and morbidity; distant metastases occur in ~40% of cases. Here, as an alternative to ionizing radiation therapy and chemotherapy and their associated side effects, we explored a new combination approach using capecitabine (CPBN) and aminolevulinate-based photodynamic therapy (PDT). We had previously developed a combination PDT approach in which 5-fluorouracil (5FU), a differentiation-promoting agent, increases the levels of protoporphyrin IX (PpIX) in cancer cells when given as a neoadjuvant prior to aminolevulinic acid (ALA). However, 5FU can be toxic when administered systemically at high levels. We reasoned that CPBN, a known chemotherapeutic for BCA and less toxic than 5FU (because CPBN is metabolized to 5FU specifically within tumor tissues), might work equally well as a PDT neoadjuvant.

Methods: Murine 4T1 BCA cells harboring a luciferase transgene were injected into breast fat pads of female nude mice. CPBN (600 mg/kg/day) was administered by oral gavage for 3 days followed by intraperitoneal ALA administration and PDT with red light (633 nm) on day 4. Tumor growth and regression were monitored in vivo using bioluminescence imaging. Histological changes in primary tumors and metastases were assessed by immunohistochemistry after necropsy.

Results: CPBN pretreatment of 4T1 tumors increased cellular differentiation, reduced proliferation, raised PpIX levels, enhanced tumor cell death, and reduced metastatic spread of 4T1 cells post-PDT, relative to vehicle-only controls.

Conclusion: The use of CPBN as a non-toxic PDT neoadjuvant for treatment of BCA represents a novel approach with significant potential for translation into the clinic.

Keywords

Photodynamic therapy / protoporphyrin IX / aminolevulinic acid / breast cancer / metastasis / cell death / 4T1 / in vivo imaging system

References

[1] Torre LA,Siegel RL,Lortet-Tieulent J.Global cancer statistics, 2012..CA Cancer J Clin2015;65:87-108

[2] Torre LA,Ward EM.Global cancer incidence and mortality rates and trends--an update..Cancer Epidemiol Biomarkers Prev2016;25:16-27

[3] Torre LA,Siegel RL,Jemal A.Global cancer in women: burden and trends..Cancer Epidemiol Biomarkers Prev2017;26:444-57

[4] Weigelt B,van ‘t Veer LJ.Breast cancer metastasis: markers and models..Nat Rev Cancer2005;5:591-602

[5] Nielsen HM,Grau C.Study of failure pattern among high-risk breast cancer patients with or without postmastectomy radiotherapy in addition to adjuvant systemic therapy: long-term results from the Danish Breast Cancer Cooperative Group DBCG 82 b and c randomized studies..J Clin Oncol2006;24:2268-75

[6] Ragaz J,Spinelli JJ,Jackson SM.Locoregional radiation therapy in patients with high-risk breast cancer receiving adjuvant chemotherapy: 20-year results of the British Columbia randomized trial..J Natl Cancer Inst2005;97:116-26

[7] Hasan T,Solban N.Photodynamic therapy of cancer.2006;7th editionHamilton, OntarioBC Decker, Inc.537-48

[8] Celli JP,Rizvi I,Samkoe KS.Imaging and photodynamic therapy: mechanisms, monitoring, and optimization..Chem Rev2010;110:2795-838 PMCID:PMC2896821

[9] Anand S,Pereira SP,Maytin EV.Biomodulatory approaches to photodynamic therapy for solid tumors..Cancer Letters2012;326:8-16 PMCID:PMC3459352

[10] Kennedy JC,Pross DC.Photodynamic therapy with endogenous protoporphyrin IX: basic principles and present clinical experience..J Photochem Photobiol B1990;6:143-8

[11] Marcus SL,Golub AL,Lundahl S.Photodynamic therapy (PDT) and photodiagnosis (PD) using endogenous photosensitization induced by 5-aminolevulinic acid (ALA): mechanisms and clinical results..J Clin Laser Med Surg1996;14:59-66

[12] Ahmad N.Mechanism of photodynamic therapy-induced cell death..Methods Enzymol2000;319:342-58

[13] Ortel B,Calzavara-Pinton P.Molecular mechanisms of photodynamic therapy..Front Biosci (Landmark Ed)2009;14:4157-72

[14] Braathen LR,Basset-Seguin N,Foley P.Guidelines on the use of photodynamic therapy for nonmelanoma skin cancer: an international consensus. International Society for Photodynamic Therapy in Dermatology, 2005..J Am Acad Dermatol2007;56:125-43

[15] Karrer S,Weiderer P,Szeimies RM.Influence of 5-aminolevulinic acid and red light on collagen metabolism of human dermal fibroblasts..J Invest Dermatol2003;120:325-31

[16] Campbell SM,Marshall R.Effect of MAL-photodynamic therapy on hypertrophic scarring..Photodiagnosis Photodyn Ther2010;7:183-8

[17] Cuenca RE,Sibata C.Breast cancer with chest wall progression: treatment with photodynamic therapy..Ann Surg Oncol2004;11:322-7

[18] Allison RR,Mang TS,Downie GH.Photodynamic therapy for chest wall recurrence from breast cancer..Photodiagnosis Photodyn Ther2004;1:157-71

[19] Liu Y,Zhang X,Zhang S.A pilot randomized clinical study of the additive treatment effect of photodynamic therapy in breast cancer patients with chest wall recurrence..J Breast Cancer2014;17:161-6 PMCID:PMC4090319

[20] Maytin EV,Riha M,Tellez A.5-Fluorouracil enhances protoporphyrin IX accumulation and lesion clearance during photodynamic therapy of actinic keratoses: a mechanism-based clinical trial..Clin Cancer Res2018;24:3026-35

[21] Sinha AK,Ortel BJ,Mai Z.Methotrexate used in combination with aminolaevulinic acid for photodynamic killing of prostate cancer cells..Br J Cancer2006;95:485-95 PMCID:PMC2360674

[22] Anand S,Hasan T,Maytin EV.Low-dose methotrexate enhances aminolevulinate-based photodynamic therapy in skin carcinoma cells in vitro and in vivo..Clin Cancer Res2009;15:3333-43 PMCID:PMC2744072

[23] Anand S,Hasan T.Vitamin D3 enhances the apoptotic response of epithelial tumors to aminolevulinate-based photodynamic therapy..Cancer Res2011;71:6040-50 PMCID:PMC3360482

[24] Anand S,Brankov N,Hasan T.Fluorouracil enhances photodynamic therapy of squamous cell carcinoma via a p53-independent mechanism that increases protoporphyrin IX levels and tumor cell death..Mol Cancer Ther2017;16:1092-101 PMCID:PMC5497500

[25] Rollakanti KR,Maytin EV.Vitamin D enhances the efficacy of photodynamic therapy in a murine model of breast cancer..Cancer Med2015;4:633-42 PMCID:PMC4430256

[26] Anand S,Maytin EV.Mechanism of differentiation-enhanced photodynamic therapy for cancer: upregulation of coproporphyrinogen oxidase by C/EBP transcription factors..Mol Cancer Ther2013;12:1638-50 PMCID:PMC3743443

[27] Miwa M,Nishida M,Ishikawa T.Design of a novel oral fluoropyrimidine carbamate, capecitabine, which generates 5-fluorouracil selectively in tumours by enzymes concentrated in human liver and cancer tissue..Eur J Cancer1998;34:1274-81

[28] Yasuno H,Yanagisawa M,Harada N.Predictive markers of capecitabine sensitivity identified from the expression profile of pyrimidine nucleoside-metabolizing enzymes..Oncol Rep2013;29:451-8

[29] Walko CM.Capecitabine: a review..Clin Ther2005;27:23-44

[30] Venturini M.Rational development of capecitabine..Eur J Cancer2002;38:3-9

[31] Tao K,Alroy J.Imagable 4T1 model for the study of late stage breast cancer..BMC Cancer2008;8:228 PMCID:PMC2529338

[32] Duan X,Guo N,Weichselbaum RR.Photodynamic therapy mediated by nontoxic core-shell nanoparticles synergizes with immune checkpoint blockade to elicit antitumor immunity and antimetastatic effect on breast cancer..J Am Chem Soc2016;138:16686-95 PMCID:PMC5667903

[33] Kolinsky K,Dugan U,Packman K.Novel regimens of capecitabine alone and combined with irinotecan and bevacizumab in colorectal cancer xenografts..Anticancer Res2009;29:91-8

[34] Toi M,Lee SJ.Capecitabine for primary breast cancer..Oncotarget2017;8:110739-40 PMCID:PMC5762279

[35] Toi M,Ohashi Y.Adjuvant capecitabine for breast cancer..N Engl J Med2017;377:791-2

[36] Reagan-Shaw S,Ahmad N.Dose translation from animal to human studies revisited..FASEB J2008;22:659-61

[37] Nair AB.A simple practice guide for dose conversion between animals and human..J Basic Clin Pharm2016;7:27-31 PMCID:PMC4804402

[38] Longley DB,Johnston PG.5-fluorouracil: mechanisms of action and clinical strategies..Nat Rev Cancer2003;3:330-8

[39] Anand S,Horst RL,Maytin EV.Combination of oral vitamin D3 with photodynamic therapy enhances tumor cell death in a murine model of cutaneous squamous cell carcinoma..Photochem Photobiol2014;90:1126-35 PMCID:PMC4186664

[40] Pulaski BA.Mouse 4T1 breast tumor model..Curr Protoc Immunol2001;

[41] Paschall AV.An orthotopic mouse model of spontaneous breast cancer metastasis..J Vis Exp2016; PMCID:PMC5091834

[42] Longo JP,Miranda-Vilela AL,de Souza LR.Prevention of distant lung metastasis after photodynamic therapy application in a breast cancer tumor model..J Biomed Nanotechnol2016;12:689-99

[43] Kim JB,Cochran E,Ang A.Non-invasive detection of a small number of bioluminescent cancer cells in vivo..PLoS One2010;5:e9364 PMCID:PMC2826408