Monitoring the Progression of Liver Fluke-Induced Cholangiocarcinoma in a Hamster Model Using Synchrotron FTIR Microspectroscopy and Focal Plane Array Infrared Imaging

Abstract

Cholangiocarcinoma (CCA) is a bile duct cancer that originates in the bile duct epithelium. Northeastern Thailand has the highest incidence of CCA, and there is a direct correlation with liver fluke (Opisthorchis viverrini) infection. The high mortality rate of CCA is a consequence of delayed diagnosis. Fourier transform infrared (FTIR) spectroscopy is a powerful technique that detects the absorbance of molecular vibrations and is perfectly suited for the interrogation of biological samples. In this study, we applied synchrotron radiation-FTIR (SR-FTIR) microspectroscopy and focal plane array (FPA-FTIR) microspectroscopy to characterize periductal fibrosis and bile duct cells progressing to CCA induced by inoculating O. viverrini metacercariae into hamsters. SR-FTIR and FPA-FTIR measurements were performed in liver sections harvested from 1-, 2-, 3-, and 6-month post-infected hamsters compared to uninfected liver tissues. Principal component analysis (PCA) of the tissue samples showed a clear discrimination among uninfected and early-stage (1 and 2 months) and cancerous-stage (3 and 6 months) tissues. The discrimination is based on intensity changes in the phosphodiester band (1081 cm −1 ), amino acid residue (∼1396 cm −1 ), and CO stretching carboxylic esters (1745 cm −1 ). Infected tissues also show definitive bands at ∼1280, 1234, and 1201 cm −1 characteristic of the collagen triplet and indicative of fibrosis. Hierarchical cluster analysis (HCA) was performed on the FPA data and showed a classification into specific cell types. Hepatocyte, fibrotic lesion, and bile duct (cancer) were classified and HCA mapping showed similar cellular distribution pattern compared to Sirius red staining. This study was also extended to less invasive sample analysis using attenuated total reflectance-FTIR (ATR-FTIR) spectroscopy. Sera from O. viverrini-infected and uninfected hamsters were analyzed using multivariate analysis, including principal component analysis (PCA), and partial least squares-discriminant analysis (PLS-DA). PCA was able to classify spectra of normal, early-stage CCA, and CCA, while the PLS-DA gave 100% accuracy for the validation. The model was established from 17 samples (11 normal, 6 cancer) in the calibration set and 9 samples in the validation set (4 normal, 2 cancer, 3 precancerous). These results indicate that FTIR-based technology is a potential tool to detect the progression of CCA, especially in the early stages of the disease.

Detection of Human Cholangiocarcinoma Markers in Serum Using Infrared Spectroscopy

“…In our previous study [18], we reported the discrimination of O. viverrini + NDMA infected from uninfected hamster sera using PCA in the fingerprint spectral region (1800-900 cm −1 ). The important spectral signatures included: (i) a band at 1745 cm −1 assigned to the lipid ester carbonyl C=O, (ii) bands at 1380-1200 cm −1 and 1034 cm −1 from collagen, (iii) a band at 1071 cm −1 from nucleic acid phosphodiester groups and iv) a band at 1153 cm −1 from polysaccharide molecules (Table 3).…”

“…In our previous study, we reported FTIR spectral discrimination between cholangiocarcinoma and normal tissues and serum samples using an animal model [18]. The discrimination was based on changes in the phosphodiester bands, amino acid, carboxylic ester and collagen molecules in tissue and serum, whereas additional bands corresponding to the amide I, II, polysaccharides and nucleic acid molecules were important in discriminating serum samples from CCA and controls [18].…”

“…In our previous study, we reported FTIR spectral discrimination between cholangiocarcinoma and normal tissues and serum samples using an animal model [18]. The discrimination was based on changes in the phosphodiester bands, amino acid, carboxylic ester and collagen molecules in tissue and serum, whereas additional bands corresponding to the amide I, II, polysaccharides and nucleic acid molecules were important in discriminating serum samples from CCA and controls [18]. In this study, we apply ATR-FTIR spectroscopy to investigate human clinical serum samples with the aim to develop a model to discriminate the spectra of CCA from healthy, hepatocellular carcinoma (HCC) and biliary disease (BD) sera using chemometrics.…”

Genomic-transcriptomic analysis reveals Syrian hamster as a superior human disease animal model

“…Of particular interest is that infection with SARS-CoV-2 in hamsters re ects some of the age demographic differences seen in COVID-19 patients [7,11]. The Syrian hamster has also been demonstrated as an ideal model for the study of cancer and cardiovascular diseases [3,12,13]. The survival rate of severe cardiovascular diseases is limited by the lack of understanding of the precise mechanism of heart failure and its often-sudden incidence, especially in young adults [14,15].…”

Probing the Anti-Cancer Potency of Sulfated Galactans on Cholangiocarcinoma Cells Using Synchrotron FTIR Microspectroscopy, Molecular Docking, and In Vitro Studies

“…In cancer research, SR-FTIR-MS has been used as a non-labeled tool for preclinical screening of anti-cancer agents and discriminating chemo-sensitive and chemo-resistant cells in different cancers [ 13 , 17 ]. Moreover, recent studies have reported the FTIR signature spectra that correlated with apoptotic CCA cells [ 18 ] and for monitoring of CCA progression in the hamster model [ 19 ].…”