Collagen analysis by second-harmonic generation microscopy predicts outcome of luminal breast cancer
Rodrigo A Natal,
José Vassallo,
Geisilene R Paiva,
Vitor B Pelegati,
Guilherme O Barbosa,
Guilherme R Mendonça,
Caroline Bondarik,
Sophie F Derchain,
Hernandes F Carvalho,
Carmen S Lima,
Carlos L Cesar,
Luís Otávio Sarian
Affiliations
Rodrigo A Natal
Laboratory of Investigative and Molecular Pathology, Center for Investigation in Pediatrics (CIPED), Faculty of Medical Sciences, State University of Campinas, Campinas, Brazil
José Vassallo
Department of Anatomic Pathology, A.C. Camargo Cancer Center, São Paulo, Brazil
Geisilene R Paiva
Laboratory of Experimental Pathology (LAPE), CAISM—Women’s Hospital, State University of Campinas, Campinas, Brazil
Vitor B Pelegati
INFABIC—National Institute of Science and Technology on Photonics Applied to Cell Biology, Campinas, Brazil
Guilherme O Barbosa
Department of Structural and Functional Biology, Institute of Biology, State University of Campinas, Campinas, Brazil
Guilherme R Mendonça
Laboratory of Investigative and Molecular Pathology, Center for Investigation in Pediatrics (CIPED), Faculty of Medical Sciences, State University of Campinas, Campinas, Brazil
Caroline Bondarik
Laboratory of Investigative and Molecular Pathology, Center for Investigation in Pediatrics (CIPED), Faculty of Medical Sciences, State University of Campinas, Campinas, Brazil
Sophie F Derchain
Department of Obstetrics and Gynecology, Faculty of Medical Sciences and CAISM—Women’s Hospital, State University of Campinas, Campinas, São Paulo, Brazil
Hernandes F Carvalho
Department of Structural and Functional Biology, Institute of Biology, State University of Campinas, Campinas, Brazil
Carmen S Lima
Oncology Section, Department of Internal Medicine, Faculty of Medical Sciences, State University of Campinas, Campinas, São Paulo, Brazil
Carlos L Cesar
INFABIC—National Institute of Science and Technology on Photonics Applied to Cell Biology, Campinas, Brazil
Luís Otávio Sarian
Department of Obstetrics and Gynecology, Faculty of Medical Sciences and CAISM—Women’s Hospital, State University of Campinas, Campinas, São Paulo, Brazil
Second-harmonic generation microscopy represents an important tool to evaluate extracellular matrix collagen structure, which undergoes changes during cancer progression. Thus, it is potentially relevant to assess breast cancer development. We propose the use of second-harmonic generation images of tumor stroma selected on hematoxylin and eosin–stained slides to evaluate the prognostic value of collagen fibers analyses in peri and intratumoral areas in patients diagnosed with invasive ductal breast carcinoma. Quantitative analyses of collagen parameters were performed using ImageJ software. These parameters presented significantly higher values in peri than in intratumoral areas. Higher intratumoral collagen uniformity was associated with high pathological stages and with the presence of axillary lymph node metastasis. In patients with immunohistochemistry-based luminal subtype, higher intratumoral collagen uniformity and quantity were independently associated with poorer relapse-free and overall survival, respectively. A multivariate response recursive partitioning model determined 12.857 and 11.894 as the best cut-offs for intratumoral collagen quantity and uniformity, respectively. These values have shown high sensitivity and specificity to differentiate distinct outcomes. Values of intratumoral collagen quantity and uniformity exceeding the cut-offs were strongly associated with poorer relapse-free and overall survival. Our findings support a promising prognostic value of quantitative evaluation of intratumoral collagen by second-harmonic generation imaging mainly in the luminal subtype breast cancer.
