Integrated Biobanking and Tumor Model Establishment of Human Colorectal Carcinoma Provides Excellent Tools for Preclinical Research

Abstract

Over the time period from 2006 to 2017, consecutive patients operated on at the University Medical Center Rostock participated in the comprehensive biobanking and tumor-modelling approach known as the HROC collection. Samples were collected using strict standard operating procedures including blood (serum and lymphocytes), tumor tissue (vital and snap frozen), and adjacent normal epithelium. Patient and tumor data including classification, molecular type, clinical outcome, and results of the model establishment are the essential pillars. Overall, 149 patient-derived xenografts with 34 primary and 35 secondary cell lines were successfully established and encompass all colorectal carcinoma anatomic sites, grading and staging types, and molecular classes. The HROC collection represents one of the largest model assortments from consecutive clinical colorectal carcinoma (CRC) cases worldwide. Statistical analysis identified a variety of clinicopathological and molecular factors associated with model success in univariate analysis. Several of them not identified before include localization, mutational status of K-Ras and B-Raf, MSI-status, and grading and staging parameters. In a multivariate analysis model, success solely correlated positively with the nodal status N1 and mutations in the genes K-Ras and B-Raf. These results imply that generating CRC tumor models on the individual patient level is worth considering especially for advanced tumor cases with a dismal prognosis.

Patient-individual cancer cell lines and tissue analysis delivers no evidence of sequences from DNA viruses in colorectal cancer cells

“…Primary aim of this study was the detection of the potentially oncogenic viruses SV40, JCV, BKV and EBV in patientindividual low passage (<P50) CRC cell lines [21]. Therefore, gDNA and cDNA from established and characterized cell lines and corresponding patient tumor (Tu) and normal (N) tissues, as well as metastases (Met) were examined by RT-PCR.…”

Patient-individual cancer cell lines and tissue analysis delivers no evidence of sequences from DNA viruses in colorectal cancer cells

“…Primary aim of this study was the detection of the potentially oncogenic viruses SV40, JCV, BKV and EBV in patient-individual low passage (<P50) CRC cell lines [21]. Therefore, gDNA and cDNA from established and characterized cell lines and corresponding patient tumor (tu) and normal (n) tissues, as well as metastases (met) were examined by RT-PCR.…”

“…In order to avoid any bias by non-tumor cells present in the tumor microenvironment, we focused on patient-individual, low passage CRC cell lines; but corresponding patient tissue samples were analyzed in addition. Using highly sensitive PCR approaches, we investigated the natural occurrence of the polyomaviruses SV40, JCV and BKV as well as the herpesvirus EBV -the most frequently discussed viruses with a potential role in CRC development [9,15,21,13,10,23,24,25,26]. First, both patientindividual CRC cell lines (n = 49) and corresponding tissue samples from patients (normal and tumor tissue, as well as liver, lung and peritoneal metastases) were analyzed for the presence of viral sequences by end-point PCR.…”

Connecting Cancer Pathways to Tumor Engines: A Stratification Tool for Colorectal Cancer Combining Human In Vitro Tissue Models with Boolean In Silico Models

“…We used HROC24 and HROC87 cells [29] cultured in DMEM/F12 medium supplemented with 10% FCS under standard culture conditions (37 • C, 5% CO 2 ).…”

“…We used MSI CRC cell lines directly derived from patient-derived xenografts (PDX) [27]. These cell lines, HROC24 and HROC87 (Hansestadt ROstock-Colon), harbor a BRAF mutation with further co-mutations: HROC24 with APC mutation and HROC87 with p53 mutation [27][28][29]. For our CRC models, we use an avascular part of a decellularized porcine gut [30][31][32][33]-called small intestine submucosa with preserved mucosa (SISmuc) [34], that is also applied for lung cancer and breast cancer models [18,19,[35][36][37].…”