scholarly journals Microsatellite Instability in Colorectal Cancer: Considerations for Molecular Diagnosis and High-Throughput Screening of Archival Tissues

2004 ◽  
Vol 50 (6) ◽  
pp. 1082-1086 ◽  
Author(s):  
Manuel Salto-Tellez ◽  
Soo Chin Lee ◽  
Lily L Chiu ◽  
Chi Kuen Lee ◽  
May Chin Yong ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Microsatellite Instability ◽  
Molecular Diagnosis ◽  
High Throughput ◽  
High Throughput Screening

scholarly journals High-throughput screening in colorectal cancer tissue-originated spheroids

2018 ◽  
Vol 110 (1) ◽  
pp. 345-355 ◽  
Author(s):  
Jumpei Kondo ◽  
Tomoya Ekawa ◽  
Hiroko Endo ◽  
Kanami Yamazaki ◽  
Norio Tanaka ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
High Throughput ◽  
High Throughput Screening ◽  
Cancer Tissue ◽  
Colorectal Cancer Tissue

scholarly journals Mapping Out Receptors: High Throughput Screening of Endothelial Monocyte Activating Polypeptide II (EMAPII) Using PRESTO-TANGO

2018 ◽  
Vol 1 (1) ◽  
Author(s):  
Nathan Lam ◽  
Bernhard Maier ◽  
Sarvesh Chelvanambi ◽  
Takashi Hato ◽  
Matthias Clauss
Keyword(s):  
Colorectal Cancer ◽  
Cardiovascular Disease ◽  
Receptor Binding ◽  
High Throughput ◽  
High Throughput Screening ◽  
Neutralizing Antibodies ◽  
Signal To Noise Ratio ◽  
Receptor System ◽  
Gpcr Activation ◽  
Cxcr3 Ligand

Background: Secreted endothelial monocyte activating polypeptide II (EMAPII/AIMP1) is a pro-apoptotic, pro-inflammatory ligand implicated in diseases such as colorectal cancer, cardiovascular disease, and emphysema. Thus, EMAPII has been shown to induce apoptosis through CXCR3 receptor binding. However, not all EMAPII functions have been attributed to CXCR3. Discovery of new receptors interacting with EMAPII could lead to development of new therapies blocking cognate ligand-receptor binding. We hypothesize the existence of at least one unknown secondary receptor for EMAPII.  Methods: The PRESTO-TANGO assay, a construct which converts G-protein coupled receptor (GPCR) ligand binding into luciferase activity measurable by luminometer, was validated using transfection with TANGO-modified CXCR3 and S1P1R as test receptors in HTLA cells. Protocols for cell transfection, adherence, and cultivation were optimized with IP10, EMAPII, and S1P as test ligands.  Results: The assay was successfully validated using several GPCR activation readouts. Binding curves were generated for S1P/S1P1 (EC50= 569nM), IP10/CXCR3 (EC50= 47.1 nM), and EMAPII/CXCR3 (EC50= 628 nM). Conditions for the PRESTO-TANGO assay were further refined for maximal signal-to-noise ratio and robust inter-assay reproducibility in preparation for high-throughput screening. We are currently testing 314 TANGO-modified GPCRs for EMAPII affinity.  Conclusion: We have validated the Tango assay for the known EMAPII-CXCR3 ligand-receptor system, a valuable tool for evaluation of anti-EMAPII therapeutics. Discovery of a novel EMAPII receptor would allow for the development of therapies including neutralizing antibodies (analogous to the PD1 receptor antibody Pembrolizumab for solid tumors) in diseases such as colorectal cancer, cardiovascular disease, and emphysema.

scholarly journals High-Throughput Screening of Combinatorial Immunotherapies with Patient-Specific In Silico Models of Metastatic Colorectal Cancer

2018 ◽  
Vol 78 (17) ◽  
pp. 5155-5163 ◽  
Author(s):  
Jakob Nikolas Kather ◽  
Pornpimol Charoentong ◽  
Meggy Suarez-Carmona ◽  
Esther Herpel ◽  
Fee Klupp ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Metastatic Colorectal Cancer ◽  
High Throughput ◽  
In Silico ◽  
High Throughput Screening ◽  
Patient Specific ◽  
In Silico Models

scholarly journals A next-generation sequencing-based strategy combining microsatellite instability and tumor mutation burden for comprehensive molecular diagnosis of advanced colorectal cancer

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jian Xiao ◽  
Wenyun Li ◽  
Yan Huang ◽  
Mengli Huang ◽  
Shanshan Li ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Microsatellite Instability ◽  
Molecular Diagnosis ◽  
Checkpoint Inhibitors ◽  
The Cancer Genome Atlas ◽  
Immune Microenvironment ◽  
Mutational Burden ◽  
Immune Signatures ◽  
Mutation Burden ◽  
Tumor Mutational Burden

Abstract Background Mismatch repair (MMR)/microsatellite instability (MSI) and tumor mutational burden (TMB) are independent biomarkers that complement each other for predicting immune checkpoint inhibitors (ICIs) efficacy. Here we aim to establish a strategy that integrates MSI and TMB determination for colorectal cancer (CRC) in one single assay. Methods Surgical or biopsy specimens retrospectively collected from CRC patients were subjected to NGS analysis. Immunohistochemistry (IHC) and polymerase chain reaction (PCR) were also used to determine MMR/MSI for those having enough tissues. The NGS-MSI method was validated against IHC and PCR. The MSI-high (MSI-H) or microsatellite stable (MSS) groups were further stratified based on tumor mutational burden, followed by validation using the The Cancer Genome Atlas (TCGA) CRC dataset. Immune microenvironment was evaluated for each subgroup be profiling the expression of immune signatures. Results Tissues from 430 CRC patients were analyzed using a 381-gene NGS panel. Alterations in KRAS, NRAS, BRAF, and HER2 occurred at a significantly higher incidence among MSI-H tumors than in MSS patients (83.6% vs. 58.4%, p = 0.0003). A subset comprising 98 tumors were tested for MSI/MMR using all three techniques, where NGS proved to be 99.0 and 93.9% concordant with PCR and IHC, respectively. Four of the 7 IHC-PCR discordant cases had low TMB (1.1–8.1 muts/Mb) and were confirmed to have been misdiagnosed by IHC. Intriguingly, 4 of the 66 MSS tumors (as determined by NGS) were defined as TMB-high (TMB-H) using a cut-off of 29 mut/Mb. Likewise, 15 of the 456 MSS tumors in the TCGA CRC cohort were also TMB-H with a cut-off of 9 muts/Mb. Expression of immune signatures across subgroups (MSS-TMB-H, MSI-H-TMB-H, and MSS-TMB-L) confirmed that the microenvironment of the MSS-TMB-H tumors was similar to that of the MSI-H-TMB-H tumors, but significantly more immune-responsive than that of the MSS-TMB-L tumors, indicating that MSI combined with TMB may be more precise than MSI alone for immune microenvironment prediction. Conclusion This study demonstrated that NGS panel-based method is both robust and tissue-efficient for comprehensive molecular diagnosis of CRC. It also underscores the importance of combining MSI and TMB information for discerning patients with different microenvironment.

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