Prognistic Value of Macrophages in CRC | Wenjing Ye
Prognostic value of CD169+ macrophages in primary colorectal cancer and synchronous or metachronous liver metastasis
Authors: Wenjing Ye (1), Andriy Trailin (1), Sergii Pavlov (1), Filip Ambrozkiewicz (1), Esraa Ali (1),
Lenka Cervenkova (1), Václav Liška (1,2)
Supervisor: Kari Hemminki (1,3)
(1) Biomedical Center, Faculty of Medicine Pilsen, Charles University (2) Department of Surgery, Faculty of Medicine in Pilsen, Charles University and University Hospital, Pilsen (3) Department of Cancer Epidemiology, German Cancer Research Center, Heidelberg, Germany.
State-of-the-Art: Macrophages in the tumor microenvironment can regulate the progression
of tumors. CD169+ macrophages are a unique type of macrophage subset that differ from M1 and M2 macrophages and are primarily expressed in secondary lymphoid organs. CD169+ macrophages have been reported to play an important role in anti-tumor immunity. Colorectal cancer (CRC) is the third most common type of cancer worldwide and the second leading cause of cancer-related deaths. The presence of distant metastases significantly reduces survival rates in CRC. The role of CD169+ macrophages in microenvironment of primary colorectal cancer and its synchronous and metachronous liver metastasis (LM) remains unclear.
Objective: We aimed at investigating distribution and prognostic value of local CD169+ macrophages
between primary colorectal cancer (pCRC) and its paired synchronous and metachronous LM.
Material and Methods: Formalin-fixed paraffin-embedded tissue sections from pCRC and synchronous (N=55) or metachronous (N=44) LM were stained by immunoperoxidase method using anti-CD169 antibodies. Densities of CD169+ macrophages were assessed in tumor center (TC), inner margin (IM), outer margin (OM) and peritumor area (PT) of pCRC and LM by using the QuPath image analysis software and assessed as prognostic variables for overall survival (OS) and disease-free survival (DFS).
Results & Discussion: The only significant difference between LM and pCRC was greater cell density in PT region of LM in synchronous group. OM and PT region showed greater CD169+ cell densities than TC and IM in pCRC and LM in both groups. Patients with synchronous LM had greater CD169+ cell densities in IM of pCRC and TC and OM of LM compared to metachronous group. High densities of CD169+ cells in the TC of pCRC in synchronous group were associated with longer OS, and their high densities in IM and PT region of pCRC in metachronous group were associated with longer DFS. Contrariwise, high densities in IM of LM in metachronous group were negatively associated with OS. Our findings highlighted unique features of tumor microenvironment between pCRC and LM and between synchronous and metachronous LM.
Conclusion: CD169+ macrophages in pCRC confer prognostic benefit for CRC patients with synchronous and metachronous LM. Contrariwise, in metachronous LM CD169+ macrophages associate with a shorter OS. We suggest CD169+ macrophages may serve as a useful prognostic marker in CRC.
Funding: AZV NW24-03-00521
Study program: Doctoral study – Experimetnal Surgery |Year of study: 3
ID: 1091
Transcriptome and miRNome in mCLM | Bhavana Hemantha Rao
Molecular insights into metachronous colorectal liver metastasis: Transcriptome and miRN ome perspectives
Authors: Bhavana Hemantha Rao (1), Veronika Boušková (1,3), Lucie Heczko (1), Petr Holý (1,3), Karolína Šeborová (1,3), Václav Liška (1,2), Ondřej Vyčítal (1,2), Ondřej Fiala (1,4), Pavel Souček (1,3), Viktor Hlaváč (1,3)
Supervisor: Viktor Hlaváč (1,2)
(1) Biomedical Center, Faculty of Medicine Pilsen, Charles University (2) Department of Surgery, Faculty of Medicine in Pilsen, Charles University and University Hospital, Pilsen (3) Toxicogenomics Unit, National Institute of Public Health, Prague, Czech Republic (4) Department of Oncology and Radiotherapeutics, Faculty of Medicine in Pilsen, Charles University and University Hospital, Pilsen
State-of-the-Art: Colorectal cancer ranks as the third most prevalent malignancy globally, with liver metastasis occurring in up to 70% of cases. Metachronous colorectal liver metastasis (mCLM), developing six months after the primary tumor’s resection, presents unique clinical challenges due to its distinct molecular profile and complex underlying mechanism. Recent advancements in transcriptomics and miRNomics have explored potential biomarkers and pathways that play a vital role in mCLM development. These studies provide insights into tumor progression, metastatic behavior, and prognostic implications.
Objective: This study aimed to investigate the transcriptome and miRNome profiles of mCLM, identifying key deregulated genes, micro RNAs (miRNAs), pathways, and their clinical relevance.
Material and Methods: Differential expression analysis was performed on 36 mCLM and adjacent non-malignant liver tissue sample pairs from RNAseq to identify deregulated genes and miRNAs. Gene set enrichment analysis and consensus molecular subtypes (CMS) classification were used to explore pathways and classify tumors. miRNA-mRNA interactions were investigated through correlation analysis, with prognostic relevance assessed by survival analysis. Validation of key interactions was performed using multiMiR database.
Results & Discussion: The transcriptomic analysis identified 1809 upregulated and 1639 downregulated genes in mCLM compared to the adjacent non-malignant liver, along with 108 upregulated and 92 downregulated miRNAs in miRNome analysis. Upregulated genes were associated to EMT, G2M checkpoints, and E2F targets, including PMEPA1, ITGA2, CDK1, CCND2, MKI67, SOX4, and S100A6. CMS classification showed that 47% of the samples were CMS2 (canonical pathway) and 22% were CMS4 (mesenchymal subtype), with distinct mutations (e.g. BRAF, KRAS, APC, TP53) and copy number variations. Key miRNA-mRNA interactions were identified, including PEA15 with hsa-miR-320b/c, TEX2/CTSO with hsa-miR-103a-3p, and PHLDA3 with hsa-miR-1304, highlighting potential regulatory mechanism in mCLM progression.
Conclusion: This study revealed holistic molecular insights into the mechanisms driving mCLM and novel miRNA-mRNA interactions with potential prognostic implications.
Funding: Funded by the Grant Agency of Charles University in Prague, programs Cooperatio “Surgical Disciplines”, no. 207043; project No: 183424; project The role of circular RNAs in chemotherapy resistance in colorectal cancer PRIMUS/25/MED/007.
Study program: Doctoral study – Experimetnal Surgery | Year of study: 3
ID: 1095
snoRNA Expression in non-viral HCC | Venkata Romana Mallela
Small nucleolar RNA expression and their prognostic values in non-viral hepatocellular carcinoma
Authors: Venkata Ramana Mallela (1), Phanindra Babu Kasi (1), Andriy Trailin (1), Dattatraya Shetti (1),
Lenka Červenková (2), Richard Pálek (2), Václav Liška (3), Kari Hemminki (4), Filip Ambrozkiewicz (1)
Supervisor: Filip Ambrozkiewicz (1)
(1) Laboratory of Translational Cancer Genomics, Biomedical Center, Faculty of Medicine in Pilsen,
Charles University (2) Laboratory of Cancer Treatment and Tissue Regeneration, Biomedical
Center, Faculty of Medicine in Pilsen, Charles University (3) Department of Surgery, Faculty of
Medicine in Pilsen, Charles University and University Hospital in Pilsen (4) German Cancer Research
Center, Heidelberg, Germany
State-of-the-Art: The absence of accurate prognostic indicators contributes to the ongoing
challenge of hepatocellular carcinoma (HCC), resulting in high mortality rates and poor prediction
of the recurrence and survival. There has been limited research exploring the relationship
between small nucleolar RNAs (snoRNAs) and HCC. Studies have previously demonstrated that
the expression of both SNORA47 and SNORD126 has been altered or dysregulated in liver cancer
tissue. In HCC, siRNA transfection suppresses SNORA47, decreasing cell invasion, proliferation,
and metastasis by modifying an epithelial-mesenchymal transition. Conversely, increased
SNORD126 levels have been associated with HCC, contributing to increased cell division.
Objective: Our aim of study is to provide a better understanding of how snoRNA expression
affects patients’ outcomes in non-viral HCC.
Material and Methods: We analyzed HCC and non-tumor adjacent tissue from 35 patients who
had undergone resection in Pilsen University hospital between 1997 and 2019. Using q-PCR, we
assessed the expression levels of specific snoRNAs, namely SNORA47 and SNORD126. Patients
were categorized into low and high expression groups based on the median expression of SNORA47
and SNORD126. We then conducted Kaplan-Meier analysis to assess the association of SNORA47 and SNORD126 expression levels with patient outcomes: time to recurrence (TTR), disease-free survival (DFS), and overall survival (OS).
Results & Discussion: SNORA47 expression was higher in tumor tissue than in non-tumor adjacent
tissue. In contrast, SNORD126 expression was lower in tumor tissues compared to non-tumor
adjacent tissue. Low expression of SNORA47 was associated with longer TTR (p = 0.03) and
DFS (p = 0.04). In univariate analysis, high expression of snoRA47 significantly correlated with
shorter DFS (HR: 2.55, 95% CI: 1.02–6.38, p = 0.05) and shorter TTR (HR: 3.85, 95% CI: 1.02–14.5,
p = 0.05). Whereas low expression of SNORD126 was associated with longer TTR (p = 0.05) but
not DFS. The combination of SNORA47-SNORD126 low expression was linked to longer TTR and
DFS (p = 0.01 and 0.02, respectively). Furthermore, no association was seen between expression
and OS. No correlations with clinical data were observed.
Conclusion: The findings suggest that SNORA47 and SNORD126 show potential as a promising
prognostic marker in non-viral HCC. The analysis revealed that the combination analysis provides
better prediction than alone for assessing the prognosis of non-viral HCC patients.
Funding: This research has received funding from the grants, and by the project National Institute
for Cancer Research—NICR (Programme EXCELES, ID Project No. LX22NPO5102), funded by the European Union—Next Generation EU.
Study program: Doctoral study – Experimetnal Surgery | Year of study: 3
ID: 1078
Whole-exome Sequencing of Colorectal Cancer | Marie Rajtmajerová
Whole-exome sequencing of primary colorectal cancer and its paired synchronous and metachronous liver metastases
Authors: Marie Rajtmajerova (1), Filip Ambrozkiewicz (1), Viktor Hlavac (1,3), Andriy Trailin (1), Lenka Cervenkova (1), Jan Bruha (1,2), Simona Susova (1,3), Pavel Vodicka (1), Ludmila Vodickova (1), Ondrej Kubecek (4), Stanislav Filip (4), Venkata Ramana Mallela (1), Wenjing Ye (1), Frantisek Zitricky (1), Vaclav Liska (1,2), Kari Hemminki (1,5)
Supervisor: Kari Hemminki (1,5)
(1) Biomedical Center, Faculty of Medicine in Pilsen, Charles University (2) Department of Surgery, Faculty of Medicine in Pilsen, Charles University and University Hospital, Pilsen (3) Toxicogenomics Unit, National Institute of Public Health, Prague (4) Department of Oncology and Radiotherapy, Faculty of Medicine and University Hospital in Hradec Kralove, Hradec Kralove (5) Department of Cancer Epidemiology, German Cancer Research Center, Heidelberg, Germany
State-of-the-Art: In spite of the fact that colorectal cancer is the second most deadly cancer worldwide, only limited number of studies focused on analysis of mutations detected in primary and metastatic tissue from the same individuals. Moreover, information about differences between tumors with synchronous metastases (which are diagnosed at the same time as primary tumor) or metachronous metastases (which are diagnosed after primary tumor removal) are even more scarce.
Objective: We aimed to investigate the differences in mutational changes associated with synchronous and metachronous course of the disease using paired patient samples of primary and metastatic cancer.
Material and Methods: We performed whole-exome sequencing of 210 FFPE patient samples of both primary colorectal cancer and liver metastases. Subsequent analyses included differential mutations, genetic interactions, copy number variation and tumor mutation burden.
Results & Discussion: APC and TP53, well known driver genes, were the most commonly mutated genes in our data set. Mutations in MPDZ gene were only detected in primary tumors of patients with metachronous metastatic disease, whereas VCAN, MTCL1, MDN1, SHROOM2, SPEG and GLI2 were significantly more prevalent in primary tumors of the synchronous group. Furthermore, different genetic interactions were present in synchronous and metachronous patients. Whilst in primary tumors of patients with synchronous metastases TP53 mutations associate with APC, in metachronous group it is mostly mutated together with NBPF11 and PRAMEF15. Paired analysis of tumor mutation burden showed positive correlation between tumor mutation burden of primary and metastatic tissue in synchronous group only.
Conclusion: Our results support the hypothesis that distinct molecular pathways underlie different time course of the disease (eg. MPDZ, VCAN, MTCL1 and other genes). However, large scale studies are required to further investigate this topic and validate our results.
Funding: This research has received funding from the Grant Agency of the Czech Republic – project number 23-05609S.
Study program: Doctoral study – Experimetnal Surgery | Year of study: 4
ID: 1114