Monitoring Breast Cancer Response to Treatment Using Stokes Shift Spectroscopy of Blood Plasma

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With the emerging trend of personalized cancer treatment, there is a need to develop noninvasive/minimally invasive techniques for treatment monitoring. In this regard, in this work fluorescence analysis of blood plasma of breast cancer patients has been used for the evaluation of response to treatment. This approach delivers information not only about the change in biochemical constituents but also about the altered metabolic pathway. Spectral deconvolution method is employed to compute the fluorescence intensity, peak wavelength, and full-width half maxima for different endogenous fluorophores. The fluorescence measurements were made on blood plasma collected from 10 normal subjects, 10 pre-treated cancer patients, and 10 post-treated patients. Besides, variations in relative concentration of tryptophan, collagen, NADH, and FAD, peak shifts and broadening of peaks are observed for tryptophan, NADH, and FAD, in blood plasma of pre-treated cancer patients indicating both biochemical and microenvironmental changes at cellular level. Further, the spectral profile of blood plasma of post-treated patients found to be similar to blood plasma of normal subjects. Linear discriminant analysis showed that pre-treated and post-treated breast cancer is discriminated with a sensitivity and specificity of 100% and 100% respectively.

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  1. 1.

    Siegel RL, Miller KD, Fedewa SA, Ahnen DJ, Meester RG, Barzi A, Jemal A (2017) Colorectal cancer statistics, 2017. CA: CA Cancer J Clin 67(3):177–193

  2. 2.

    Harris L, Fritsche H, Mennel R, Norton L, Ravdin P, Taube S, Somerfield MR, Hayes DF, Bast RC Jr (2007) American Society of Clinical Oncology 2007 update of recommendations for the use of tumor markers in breast cancer. J Clin Oncol 25(33):5287–5312

  3. 3.

    Erdi YE (2012) Limits of tumor detectability in nuclear medicine and PET. Mol Imaging Radionucl Ther 21(1):23

  4. 4.

    McCormack DR, Walsh AJ, Sit W, Arteaga CL, Chen J, Cook RS, Skala MC (2014) In vivo hyperspectral imaging of microvessel response to trastuzumab treatment in breast cancer xenografts. Biomed Opt Express 5(7):2247–2261

  5. 5.

    Tromberg BJ, Zhang Z, Leproux A, O'Sullivan TD, Cerussi AE, Carpenter PM, Mehta RS, Roblyer D, Yang W, Paulsen KD, Pogue BW (2016) Predicting responses to neoadjuvant chemotherapy in breast cancer: ACRIN 6691 trial of diffuse optical spectroscopic imaging. Cancer Res 76(20):5933–5944

  6. 6.

    Vidyasagar MS, Maheedhar K, Vadhiraja BM, Fernendes DJ, Kartha VB, Krishna CM (2008) Prediction of radiotherapy response in cervix cancer by Raman spectroscopy: a pilot study. Biopolymers 89(6):530–537

  7. 7.

    Tsuyuki S, Yamaguchi A, Kawata Y, Kawaguchi K (2015) Assessing the effects of neoadjuvant chemotherapy on lymphatic pathways to sentinel lymph nodes in cases of breast cancer: usefulness of the indocyanine green-fluorescence method. Breast 24(3):298–301

  8. 8.

    Pu Y, Tang GC, Wang WB, Savage HE, Schantz SP, Alfano RR (2011) Native fluorescence spectroscopic evaluation of chemotherapeutic effects on malignant cells using nonnegative matrix factorization analysis. Technol Cancer Res Treat 10(2):113–120

  9. 9.

    Pu Y, Tang G, Wang W, Savage HE, Schantz SP, Alfano RR (2012) Chemotherapeutic effects on breast malignant cells evaluated by native fluorescence spectroscopy. In: Biomedical optics JM3A-45

  10. 10.

    Chithra K, Vijayaraghavan S, Prakasarao A, Singaravelu G (2017) Study of anti-cancer effects of chemotherapeutic agents and radiotherapy in breast cancer patients using fluorescence spectroscopy. In: Optical biopsy XV: toward real-time spectroscopic imaging and diagnosis, vol 10060, p 100600L

  11. 11.

    Sivabalan S, Vedeswari CP, Jayachandran S, Koteeswaran D, Pravda C, Aruna P, Ganesan S (2010) In vivo native fluorescence spectroscopy and nicotinamide adinine dinucleotide/flavin adenine dinucleotide reduction and oxidation states of oral submucous fibrosis for chemopreventive drug monitoring. J Biomed Opt 15(1):017010

  12. 12.

    Han SH, Song TK (2015) In vivo fluorescence spectroscopic monitoring of radiotherapy in cancer treatment. Int J Cancer Ther Oncol 3(1):03013

  13. 13.

    Brown JQ, Bydlon TM, Richards LM, Yu B, Kennedy SA, Geradts J, Wilke LG, Junker MK, Gallagher J, Barry WT, Ramanujam N (2010) Optical assesssment of tumor resection margins in the breast. IEEE J Sel Top Quantum Electron 16(3):530–544

  14. 14.

    Alfano RR, Tata DB, Cordero J, Tomashefsky P, Longo F, Alfano M (1984) Laser induced fluorescence spectroscopy from native cancerous and normal tissue. IEEE J Quantum Electron 20(12):1507–1511

  15. 15.

    Zhadin NN, Yang Y, Ganesan S, Ockman N, Alfano RR (1996) Enhancement of the fluorescence cancer diagnostic method of tissues using diffuse reflectance and the analysis of oxygenation state. In: Advances in laser and light spectroscopy to diagnose cancer and other diseases III: optical biopsy, vol 2679, pp 142–149

  16. 16.

    Ganesan S, Sacks PG, Yang Y, Katz A, Al-Rawi M, Savage HE, Schantz SP, Alfano RR (1998) Native fluorescence spectroscopy of normal and malignant epithelial cells. Cancer Biochem Biophys 16(4):365–373

  17. 17.

    Liu Q, Grant G, Vo-Dinh T (2010) Investigation of synchronous fluorescence method in multicomponent analysis in tissue. IEEE J Sel Top Quantum Electron 16(4):927–940

  18. 18.

    Majumder SK (2000) Synchronous luminescence spectroscopy for oral cancer diagnosis. Lasers Life Sci 9:143–152

  19. 19.

    Vengadesan N, Anbupalam T, Hemamalini S, Ebenezar J, Muthvelu K, Koteeswaran D, Aruna PR, Ganesan S (2002) Characterization of cervical normal and abnormal tissues by synchronous luminescence spectroscopy. In Optical Biopsy IV 4613:13–18

  20. 20.

    Alfano RR, Yang Y (2003) Stokes shift emission spectroscopy of human tissue and key biomolecules. IEEE J Sel Top Quantum Electron 9(2):148–153

  21. 21.

    Dramicanin T, Dramicanin MD, Dimitrijevic B, Jokanovic V, Lukic S (2006) Discrimination between normal and malignant breast tissues by synchronous luminescence spectroscopy. Acta Chim Slov 53(4):444

  22. 22.

    Ebenezar J, Aruna P, Ganesan S (2010) Synchronous fluorescence spectroscopy for the detection and characterization of cervical cancers in vitro. Photochem Photobiol 86(1):77–86

  23. 23.

    Pu Y, Wang W, Yang Y, Alfano RR (2013) Stokes shift spectroscopic analysis of multifluorophores for human cancer detection in breast and prostate tissues. J Biomed Opt 18(1):017005

  24. 24.

    Muthuvelu K, Shanmugam S, Koteeswaran D, Srinivasan S, Venkatesan P, Aruna P, Ganesan S (2011) Synchronous luminescence spectroscopic characterization of blood elements of normal and patients with cervical cancer. In: Optical biopsy IX, vol 7895, p 78950F International Society for Optics and Photonics

  25. 25.

    Rajasekaran R, Aruna P, Koteeswaran D, Baludavid M, Ganesan S (2014) Synchronous luminescence spectroscopic characterization of urine of normal subjects and cancer patients. J Fluoresc 24(4):1199–1205

  26. 26.

    Kumar P, Ashutosh K, Pradhan A (2018) Comparative study between diagnostic mediums: human tissue and saliva for oral cancer detection using stokes shift spectroscopy. In: Optics in Health Care and Biomedical Optics VIII, vol 10820, p 108200L International Society for Optics and Photonics

  27. 27.

    Yuvaraj M, Aruna P, Koteeswaran D, Muthuvelu K, Ganesan S (2018) UV-native fluorescence steady and excited state kinetics of salivary protein of normal subjects, oral premalignant and malignant conditions. J Lumin 196:236–243

  28. 28.

    Xu X, Meng J, Hou S, Ma H, Wang D (1988) The characteristic fluorescence of the serum of cancer patients. J Lumin 40:219–220

  29. 29.

    Hubmann MR, Leiner MJ, Schaur RJ (1990) Ultraviolet fluorescence of human sera: I. sources of characteristic differences in the ultraviolet fluorescence spectra of sera from normal and cancer-bearing humans. Clin Chem 36(11):1880–1883

  30. 30.

    Madhuri S, Vengadesan N, Aruna P, Koteeswaran D, Venkatesan P, Ganesan S (2003) Native fluorescence spectroscopy of blood plasma in the characterization of Oral malignancy. Photochem Photobiol 78(2):197–204

  31. 31.

    Chauhan P, Yadav R, Kaushal V, Beniwal P (2016) Evaluation of serum biochemical profile of breast cancer patients. Int J Med Res Health Sci 5(7):1

  32. 32.

    Gnanatheepam E, Kanniyappan U, Dornadula K, Prakasarao A, Singaravelu G (2019) Synchronous luminescence spectroscopy as a tool in the discrimination and characterization of Oral Cancer tissue. J Fluoresc:1–7

  33. 33.

    Yang Y, Katz A, Celmer EJ, Zurawska-Szczepaniak M, Alfano RR (1997) Fundamental differences of excitation spectrum between malignant and benign breast tissues. Photochem Photobiol 66(4):518–522

  34. 34.

    Cascino A, Cangiano C, Ceci F, Franchi F, Mineo T, Mulieri M, Muscaritoli M, Rossi FF (1991) Increased plasma free tryptophan levels in human cancer: a tumor related effect? Anticancer Res 11(3):1313–1316

  35. 35.

    Miyagi Y, Higashiyama M, Gochi A, Akaike M, Ishikawa T, Miura T, Saruki N, Bando E, Kimura H, Imamura F, Moriyama M (2011) Plasma free amino acid profiling of five types of cancer patients and its application for early detection. PLoS One 6(9):e24143

  36. 36.

    Zhang L, Pu Y, Xue J, Pratavieira S, Xu B, Achilefu S, Alfano RR (2014) Tryptophan as the fingerprint for distinguishing aggressiveness among breast cancer cell lines using native fluorescence spectroscopy. J Biomed Opt 19(3):037005

  37. 37.

    Brisson BK, Mauldin EA, Lei W, Vogel LK, Power AM, Lo A, Dopkin D, Khanna C, Wells RG, Puré E, Volk SW (2015) Type III collagen directs stromal organization and limits metastasis in a murine model of breast cancer. Am J Pathol 185(5):1471–1486

  38. 38.

    Schomacker KT, Frisoli JK, Compton CC, Flotte TJ, Richter JM, Nishioka NS, Deutsch TF (1992) Ultraviolet laser-induced fluorescence of colonic tissue: basic biology and diagnostic potential. Lasers Surg Med 12(1):63–78

  39. 39.

    Pandey K, Pradhan A, Agarwal A, Bhagoliwal A, Agarwal N (2012) Fluorescence spectroscopy: a new approach in cervical cancer. J Obstet Gynaecol India 62(4):432–436

  40. 40.

    Mazouni C, Arun B, Andre F, Ayers M, Krishnamurthy S, Wang B, Hortobagyi GN, Buzdar AU, Pusztai L (2008) Collagen IV levels are elevated in the serum of patients with primary breast cancer compared to healthy volunteers. Br J Cancer 99(1):68

  41. 41.

    Miller JA, Pappan K, Thompson PA, Want EJ, Siskos AP, Keun HC, Wulff J, Hu C, Lang JE, Chow HHS (2015) Plasma metabolomic profiles of breast cancer patients after short-term limonene intervention. Cancer Prev Res 8(1):86–93

  42. 42.

    Chance B, Schoener B, Oshino R, Itshak F, Nakase Y (1979) Oxidation-reduction ratio studies of mitochondria in freeze-trapped samples. NADH and flavoprotein fluorescence signals. J Biol Chem 254(11):4764–4771

  43. 43.

    Pradhan A, Pal P, Durocher G, Villeneuve L, Balassy A, Babai F, Gaboury L, Blanchard L (1995) Steady state and time-resolved fluorescence properties of metastatic and non-metastatic malignant cells from different species. J Photochem Photobiol B 31(3):101–112

  44. 44.

    Glassman WS, Liu CH, Tang GC, Lubicz S, Alfano R (1992) Ultraviolet excited fluorescence spectra from non-malignant and malignant tissues of the gynecological tract. Lasers Life Sci 5(1–2):49–58

  45. 45.

    Ostrander JH, McMahon CM, Lem S, Millon SR, Brown JQ, Seewaldt VL, Ramanujam N (2010) Optical redox ratio differentiates breast cancer cell lines based on estrogen receptor status. Cancer Res 70(11):4759–4766

  46. 46.

    Wojcieszyńska D, Hupert-Kocurek K, Guzik U (2012) Flavin-dependent enzymes in cancer prevention. Int J Mol Sci 13(12):16751–16768

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Authors gratefully acknowledge DAE-BRNS (No.:2009/34/38/BRNS) for funding this work.

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Correspondence to Singaravelu Ganesan.

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Chithra, K., Aruna, P., Einstein, G. et al. Monitoring Breast Cancer Response to Treatment Using Stokes Shift Spectroscopy of Blood Plasma. J Fluoresc 29, 803–812 (2019) doi:10.1007/s10895-019-02399-9

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  • Stokes shift spectroscopy
  • Blood plasma
  • Breast cancer
  • Spectral deconvolution method
  • Treatment monitoring