Second harmonic generation for collagen I characterization in rectal cancer patients with and without preoperative radiotherapy

Abstract. Rectal cancer is treated with preoperative radiotherapy (RT) to downstage the tumor, reduce local recurrence, and improve patient survival. Still, the treatment outcome varies significantly and new biomarkers are desired. Collagen I (Col-I) is a potential biomarker, which can be visualized label-free by second harmonic generation (SHG). Here, we used SHG to identify Col-I changes induced by RT in surgical tissue, with the aim to evaluate the clinical significance of RT-induced Col-I changes. First, we established a procedure for quantitative evaluation of Col-I by SHG in CDX2-stained tissue sections. Next, we evaluated Col-I properties in material from 31 non-RT and 29 RT rectal cancer patients. We discovered that the Col-I intensity and anisotropy were higher in the tumor invasive margin than in the inner tumor and normal mucosa, and RT increased and decreased the intensity in inner tumor and normal mucosa, respectively. Furthermore, higher Col-I intensity in the inner tumor was related to increased distant recurrence in the non-RT group but to longer survival in the RT group. In conclusion, we present a new application of SHG for quantitative analysis of Col-I in surgical material, and the first data suggest Col-I intensity as a putative prognostic biomarker in rectal cancer.

[1]  Jürgen Popp,et al.  From molecular structure to tissue architecture: collagen organization probed by SHG microscopy , 2013, Journal of biophotonics.

[2]  J. Scholefield,et al.  Increased type-IV collagenase (MMP-2 and MMP-9) activity following preoperative radiotherapy in rectal cancer , 2000, British Journal of Cancer.

[3]  L Påhlman,et al.  Improved survival with preoperative radiotherapy in resectable rectal cancer. , 1997, The New England journal of medicine.

[4]  A. Enejder,et al.  X-radiation enhances the collagen type I strap formation and migration potentials of colon cancer cells , 2016, Oncotarget.

[5]  A. Jemal,et al.  Global Cancer Statistics , 2011 .

[6]  Sergey Plotnikov,et al.  Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure , 2012, Nature Protocols.

[7]  Paolo P. Provenzano,et al.  Collagen reorganization at the tumor-stromal interface facilitates local invasion , 2006, BMC medicine.

[8]  Z. Werb,et al.  The extracellular matrix: A dynamic niche in cancer progression , 2012, The Journal of cell biology.

[9]  Paul Campagnola,et al.  Second Harmonic Generation Imaging Distinguishes Both High-Grade Dysplasia and Cancer from Normal Colonic Mucosa , 2014, Digestive Diseases and Sciences.

[10]  J. Yarnold,et al.  Pathogenetic mechanisms in radiation fibrosis. , 2010, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[11]  Yuming Liu,et al.  Computational segmentation of collagen fibers from second-harmonic generation images of breast cancer , 2014, Journal of biomedical optics.

[12]  Raghu Kalluri,et al.  Fibroblasts in cancer , 2006, Nature Reviews Cancer.

[13]  Mei Xu,et al.  CDX2 as a marker for intestinal differentiation: Its utility and limitations. , 2011, World journal of gastrointestinal surgery.

[14]  Viktor H. Koelzer,et al.  The Tumor Border Configuration of Colorectal Cancer as a Histomorphological Prognostic Indicator , 2014, Front. Oncol..

[15]  Dominique Ardail,et al.  Radiation-enhanced cell migration/invasion process: a review. , 2014, Critical reviews in oncology/hematology.

[16]  Hiroshi Shimizu,et al.  Second harmonic generation and multiphoton microscopic detection of collagen without the need for species specific antibodies. , 2011, Burns : journal of the International Society for Burn Injuries.

[17]  R. Palmqvist,et al.  Liver-metastatic potential of colorectal cancer is related to the stromal composition of the tumour. , 2012, Anticancer research.

[18]  Hernandes F. Carvalho,et al.  Colon Adenocarcinoma Diagnosis in Human Samples by Multicontrast Nonlinear Optical Microscopy of Hematoxylin and Eosin Stained Histological Sections , 2014 .

[19]  Arezki Boudaoud,et al.  FibrilTool, an ImageJ plug-in to quantify fibrillar structures in raw microscopy images , 2014, Nature Protocols.

[20]  M. O’Neil,et al.  Histopathology of Colorectal Cancer after Neoadjuvant Chemoradiation Therapy , 2009 .

[21]  Paolo P. Provenzano,et al.  Aligned Collagen Is a Prognostic Signature for Survival in Human Breast Carcinoma Address Reprint Requests to See Related Commentary on Page 966 , 2022 .

[22]  Yarong Wang,et al.  Direct visualization of macrophage-assisted tumor cell intravasation in mammary tumors. , 2007, Cancer research.

[23]  S. Huerta,et al.  Mechanisms of resistance to ionizing radiation in rectal cancer , 2009, Expert review of molecular diagnostics.

[24]  L. Påhlman.  Indications for neoadjuvant long-term radiotherapy. , 2005, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.

[25]  C. Mathers,et al.  Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012 , 2015, International journal of cancer.

[26]  An Hendrix,et al.  The extracellular matrix regulates cancer progression and therapy response: implications for prognosis and treatment. , 2009, Current pharmaceutical design.

[27]  Rieken,et al.  [Preoperative radiotherapy combined with total mesorectal excision for resectable rectal cancer]. , 2001, Strahlentherapie und Onkologie : Organ der Deutschen Rontgengesellschaft ... [et al].

[28]  Yan Li,et al.  Collagen as a double-edged sword in tumor progression , 2013, Tumor Biology.

[29]  V. Centonze,et al.  Multiphoton excitation provides optical sections from deeper within scattering specimens than confocal imaging. , 1998, Biophysical journal.

[30]  Xiao-Feng Sun,et al.  Stromal staining for PINCH is an independent prognostic indicator in colorectal cancer. , 2004, Neoplasia.