Bridging the Species Gap: Morphological and Molecular Comparison of Feline and Human Intestinal Carcinomas

Limited availability of in vivo experimental models for invasive colorectal cancer (CRC) including metastasis and high tumor budding activity is a major problem in colorectal cancer research. In order to compare feline and human intestinal carcinomas, tumors of 49 cats were histologically subtyped, graded and further characterized according to the human WHO classification. Subsequently, feline tumors were compared to a cohort of 1004 human CRC cases. Feline intestinal tumors closely resembled the human phenotype on a histomorphological level. In both species, adenocarcinoma not otherwise specified (ANOS) was the most common WHO subtype. In cats, the second most common subtype of the colon (36.4%), serrated adenocarcinoma (SAC), was overrepresented compared to human CRC (8.7%). Mucinous adenocarcinoma (MAC) was the second most common subtype of the small intestine (12.5%). Intriguingly, feline carcinomas, particularly small intestinal, were generally of high tumor budding (Bd) status (Bd3), which is designated an independent prognostic key factor in human CRC. We also investigated the relevance of feline CTNNB1 exon 2 alterations by Sanger sequencing. In four cases of feline colonic malignancies (3 ANOS, 1 SAC), somatic missense mutations of feline CTNNB1 (p.D32G, p.D32N, p.G34R, and p.S37F) were detected, indicating that mutational alterations of the WNT/β-catenin signaling pathway potentially play an essential role in feline intestinal tumorigenesis comparable to humans and dogs. These results indicate that spontaneous intestinal tumors of cats constitute a useful but so far underutilized model for human CRC. Our study provides a solid foundation for advanced comparative oncology studies and emphasizes the need for further (molecular) characterization of feline intestinal carcinomas.

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The DDX39B/FUT3/TGFβR-I Axis Promotes Tumor Metastasis and EMT in Colorectal Cancer

10.21203/rs.3.rs-42529/v1 ◽

2020 ◽

Author(s):

Chengcheng He ◽

Aimin Li ◽

Qiuhua Lai ◽

Jian Ding ◽

Qun Yan ◽

...

Keyword(s):

Colorectal Cancer ◽

Nuclear Export ◽

Rna Binding ◽

Mrna Splicing ◽

Migration And Invasion ◽

Rna Seq ◽

Exon 2 ◽

Mesenchymal Transition ◽

Qrt Pcr

Abstract Background: DDX39B is a member of the DEAD box (DDX) RNA helicase family required for nearly all cellular RNA metabolism processes. The exact role and potential molecular mechanism of DDX39B in the progression of human colorectal cancer (CRC) remain to be investigated. Methods: Western blotting and quantitative real-time PCR (qRT-PCR) were conducted to detect the expression of DDX39B in CRC tissues and cell lines. Transwell and wound healing assays were conducted to assess the migration and invasion ability of CRC cells with DDX39B overexpressed or silencing. Orthotopic transplantation model of nude mice was performed to validate CRC metastasis in vivo. RNA sequencing (RNA-seq) and RNA binding protein immunoprecipitation (RIP) assay verified the direct regulation of DDX39B on the splicing and nuclear export of FUT3 mRNA, cytoplasmic and nuclear RNA isolation confirmed the nuclear export effect of DDX39B on FUT3. qRT-PCR was conducted to quantify FUT3 splicing variants. Lectin blotting was conducted to evaluate the fucosylation level of TGFβR-I.Results: In the present study, we demonstrate that DDX39B expression was higher in CRC tissues than in adjacent normal tissues. Gain- and loss- of- function assays revealed that DDX39B facilitated the metastasis of CRC in vivo and in vitro. Mechanistically, RNA-seq and RIP showed that DDX39B upregulated FUT3 expression by binding the first exon of FUT3 mRNA, which promote the mRNA splicing and export of FUT3. RNA-seq results and qRT-PCR showed that overexpression of DDX39B may favor the longer FUT3 mRNA products that contain the complete and longer exon 2, suggesting an alternative splicing of FUT3. Upregulation of FUT3 accelerated the fucosylation of TGFβR-I, thus activating the TGFβ/SMAD signaling pathway, eventually driving the epithelial-mesenchymal transition (EMT) program and contributing to CRC progression. Conclusions: Our finding demonstrated for the first time that the DDX39B/FUT3/TGFβR-I axis promotes the progression of CRC. These findings not only provide new insight into the role of DDX39B in mRNA splicing and export and tumorigenesis, but also shed light on the effect of aberrant fucosylation on CRC progression.

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Cell Line Derived Xenograft Mouse Models Are a Suitable in vivo Model for Studying Tumor Budding in Colorectal Cancer

Frontiers in Medicine ◽

10.3389/fmed.2019.00139 ◽

2019 ◽

Vol 6 ◽

Cited By ~ 5

Author(s):

Laurent M. C. Georges ◽

Olivier De Wever ◽

José A. Galván ◽

Heather Dawson ◽

Alessandro Lugli ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Line ◽

Mouse Models ◽

Tumor Budding ◽

In Vivo Model

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The Ultrastructural Analysis of Human Colorectal Cancer Stem Cell-Derived Spheroids and Their Mouse Xenograft Shows That the Same Cells Types Have Different Ratios

Biology ◽

10.3390/biology10090929 ◽

2021 ◽

Vol 10 (9) ◽

pp. 929

Author(s):

Michela Relucenti ◽

Federica Francescangeli ◽

Maria Laura De De Angelis ◽

Vito D'Andrea ◽

Selenia Miglietta ◽

...

Keyword(s):

Colorectal Cancer ◽

Stem Cell ◽

Cancer Cells ◽

Drug Testing ◽

Flow Cytometric Analysis ◽

Experimental Models ◽

Stem Cell Marker ◽

Multicellular Spheroids

Spheroids from primary colorectal cancer cells and their mice xenografts have emerged as useful preclinical models for cancer research as they replicate tumor features more faithfully as compared to cell lines. While 3D models provide a reliable system for drug discovery and testing, their structural complexity represents a challenge and their structure-function relationships are only partly understood. Here, we present a comparative ultrastructural and flow citometric analysis of patient colorectal cancer-derived spheroids and their mice xenografts. Ultrastructural observations highlighted that multicellular spheroids and their xenografts contain the same cancer cell types but with different ratios, specifically multicellular spheroids were enriched in cells with a stem-like phenotype, while xenografts had an increased amount of lipid droplets-containing cells. The flow cytometric analysis for stem cell marker and activity showed enrichment of stem-like cells presence and activity in spheroids while xenografts had the inverse response. Our results evidence the effects on cancer cells of different in vitro and in vivo microenvironments. Those differences have to be paid into account in designing innovative experimental models for personalized drug testing.

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Different hotspot p53 mutants exert distinct phenotypes and predict outcome of colorectal cancer patients

10.21203/rs.3.rs-523301/v1 ◽

2021 ◽

Author(s):

Ori Hassin ◽

Nishanth Belugali Nataraj ◽

Michal Shreberk-Shaked ◽

Yael Aylon ◽

Rona Yaeger ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Progression ◽

Regulatory Elements ◽

Migration And Invasion ◽

Missense Mutations ◽

Oncogenic Signaling ◽

Transcriptional Signature ◽

Colorectal Cancer Patients

Abstract Colorectal cancer (CRC) is the third most common cancer worldwide. The TP53 gene is mutated in approximately 60% of all CRC cases. Sporadic CRC is characterized by high prevalence of TP53 hotspot missense mutations. In particular, over 20 percent of all TP53-mutated CRC tumors carry either the p53R175H structural mutant or the p53R273H DNA contact mutant. Importantly, clinical data analysis suggests that CRC tumors harboring p53 R273 mutations are more prone to progress to metastatic disease than those with R175 mutations, in association with decreased survival. By combining in vitro CRC cell line models and human CRC data mining, we identified a unique transcriptional signature orchestrated by p53R273H, implicating activation of oncogenic signaling pathways and predicting worse patient outcome. Concordantly, p53R273H selectively promotes rapid CRC cell spreading, migration and invasion in vitro and metastasis in vivo. Mechanistically, the transcriptional output of p53R273H is associated with, and presumably driven by, its preferential binding to regulatory elements of R273 signature genes. Together, this demonstrates that different TP53 missense mutations contribute differently to cancer progression, and that p53R273H possesses distinct gain-of-function activities in CRC that bear on disease course and possibly on patient management strategy. Given that practically all current analytical cancer gene panels include TP53, elucidation of the differential impact of distinct TP53 mutations on disease features is expected to make information on TP53 mutations more actionable and holds potential for better precision-based medicine.

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Selection of a Malignant Subpopulation from a Colorectal Cancer Cell Line

10.1101/578823 ◽

2019 ◽

Author(s):

Pei-Lun Lai ◽

Ting-Chun Chen ◽

Chun-Yen Feng ◽

Hsuan Lin ◽

Ng Wu ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Line ◽

Prognostic Biomarker ◽

Differentially Expressed Gene ◽

Cancer Cell Line ◽

Experimental Models ◽

Colorectal Cancer Cell Line ◽

Selection Of

AbstractColorectal cancer (CRC) is a leading cause of death from cancer worldwide. Thus, there is an emerging need for new experimental models that allow identification and validation of biomarkers for CRC-specific progression. In this study, we propose a repeated sphere-forming assay as a strategy to select a malignant subpopulation from a CRC line, HCT116. We validated our assay by confirming that three canonical stemness markers, Nanog, Oct4, and Lgr5, were up-regulated in the sphere state at every generation of the selection assay. The resulting line, after eight rounds of selection, exhibited an increased sphere-forming capacityin vitroand tumorgenicityin vivo. Furthermore, dipeptidase 1 (DPEP1) was identified as the major differentially expressed gene in the selected clone, and depletion of DPEP1 suppressed the elevated sphere-forming capacityin vitroand tumorgenicityin vivo. Overall, we have established an experimental strategy for the isolation of a malignant subpopulation from a CRC cell line. Results from our model also suggested that DPEP1 can serve as a promising prognostic biomarker for CRC.

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Pro-protein convertase subtilisin/kexin type 9 promotes intestinal tumor development by activating Janus kinase 2/signal transducer and activator of transcription 3/SOCS3 signaling in ApcMin/+ mice

International Journal of Immunopathology and Pharmacology ◽

10.1177/20587384211038345 ◽

2021 ◽

Vol 35 ◽

pp. 205873842110383

Author(s):

Kai Yang ◽

Jie Zhu ◽

Huan-hua Luo ◽

Shu-wen Yu ◽

Lu Wang

Keyword(s):

Colorectal Cancer ◽

Density Lipoprotein ◽

Janus Kinase ◽

Cytokine Signaling ◽

Intestinal Tumor ◽

Signal Transducer ◽

Janus Kinase 2 ◽

Intestinal Tumors ◽

Transcription 3

Introduction Pro-protein convertase subtilisin/kexin type 9 (PCSK9) regulates lipoprotein homeostasis in humans. Evolocumab is a selective PCSK9 inhibitor that can reduce low-density lipoprotein cholesterol (LDLC) level and decrease hypercholesterolemia. The current study aimed to explore whether PCSK9 increases the risk of colorectal cancer. Methods First, we utilized the classic intestinal tumor ApcMin/+ mouse model and PCSK9 knock-in (KI) mice to establish ApcMin/+PCSK9(KI) mice. Then, we investigated the effect of PCSK9 overexpression in ApcMin/+PCSK9(KI) mice and PCSK9 inhibition using evolocumab on the progression of intestinal tumors in vivo by hematoxylin and eosin (HE) staining, Western blot, and immunohistochemistry (IHC) assay. Results ApcMin/+PCSK9(KI) mice had higher numbers and larger sizes of adenomas, with 83.3% of these mice developing adenocarcinoma (vs. 16.7% of ApcMin/+ mice). However, treatment with evolocumab reduced the number and size of adenomas and prevented the development of adenocarcinomas in ApcMin/+ mice. PCSK9 overexpression reduced tumor cell apoptosis, the Bax/bcl-2 ratio, and the levels of cytokine signaling 3 protein (SOCS3) suppressors, but activated Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling in intestinal tumors. In contrast, evolocumab treatment had the opposite effect on ApcMin/+mice. Conclusion PCSK9 might act as an oncogene or have an oncogenic role in the development and progression of colorectal cancer in vivo via activation of JAK2/STAT3/SOCS3 signaling.

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Mutation Spectrum in Patients with Wiskott-Aldrich Syndrome and X-linked Thrombocytopenia: Identification of Twelve Different Mutations in the WASP Gene

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650318 ◽

1996 ◽

Vol 75 (04) ◽

pp. 546-550 ◽

Cited By ~ 21

Author(s):

Marianne Schwartz ◽

Albert Békássy ◽

Mikael Donnér ◽

Thomas Hertel ◽

Stefan Hreidarson ◽

...

Keyword(s):

Base Pair ◽

Hot Spot ◽

Missense Mutations ◽

Nucleotide Substitutions ◽

Exon 2 ◽

Wiskott Aldrich Syndrome ◽

Base Pair Deletion ◽

Reliable Diagnosis ◽

Wasp Gene ◽

Detection Of Mutations

SummaryTwelve different mutations in the WASP gene were found in twelve unrelated families with Wiskott-Aldrich syndrome (WAS) or X-linked thrombocytopenia (XLT). Four frameshift, one splice, one nonsense mutation, and one 18-base-pair deletion were detected in seven patients with WAS. Only missense mutations were found in five patients diagnosed as having XLT. One of the nucleotide substitutions in exon 2 (codon 86) results in an Arg to Cys replacement. Two other nucleotide substitutions in this codon, R86L and R86H, have been reported previously, both giving rise to typical WAS symptoms, indicating a mutational hot spot in this codon. The finding of mutations in the WASP gene in both WAS and XLT gives further evidence of these syndromes being allelic. The relatively small size of the WASP gene facilitates the detection of mutations and a reliable diagnosis of both carriers and affected fetuses in families with WAS or XLT.

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Neutralization of a Low Molecular Weight Heparin (LHN-1) and Conventional Heparin by Protamine Sulfate in Rats

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661675 ◽

1986 ◽

Vol 56 (03) ◽

pp. 318-322 ◽

Cited By ~ 18

Author(s):

V Diness ◽

P B Østergaard

Keyword(s):

Molecular Weight ◽

Low Molecular Weight Heparin ◽

Thrombus Formation ◽

Experimental Models ◽

Protamine Sulfate ◽

Low Molecular Weight ◽

Antithrombotic Effect ◽

Higher Doses

SummaryThe neutralization of a low molecular weight heparin (LHN-1) and conventional heparin (CH) by protamine sulfate has been studied in vitro and in vivo. In vitro, the APTT activity of CH was completely neutralized in parallel with the anti-Xa activity. The APTT activity of LHN-1 was almost completely neutralized in a way similar to the APTT activity of CH, whereas the anti-Xa activity of LHN-1 was only partially neutralized.In vivo, CH 3 mg/kg and LHN-1 7.2 mg/kg was given intravenously in rats. The APTT and anti-Xa activities, after neutralization by protamine sulfate in vivo, were similar to the results in vitro. In CH treated rats no haemorrhagic effect in the rat tail bleeding test and no antithrombotic effect in the rat stasis model was found at a protamine sulfate to heparin ratio of about 1, which neutralized APTT and anti-Xa activities. In LHN-1 treated rats the haemorrhagic effect was neutralized when APTT was close to normal whereas higher doses of protamine sulfate were required for neutralization of the antithrombotic effect. This probably reflects the fact that in most experimental models higher doses of heparin are needed to induce bleeding than to prevent thrombus formation. Our results demonstrate that even if complete neutralization of APTT and anti-Xa activities were not seen in LHN-1 treated rats, the in vivo effects of LHN-1 could be neutralized as efficiently as those of conventional heparin. The large fall in blood pressure caused by high doses of protamine sulfate alone was prevented by the prior injection of LHN-1.

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Antimicrobial peptides for the treatment of pulmonary tuberculosis, allies or foes?

Current Pharmaceutical Design ◽

10.2174/1381612824666180327162357 ◽

2018 ◽

Vol 24 (10) ◽

pp. 1138-1147

Author(s):

Bruno Rivas-Santiago ◽

Flor Torres-Juarez

Keyword(s):

Pulmonary Tuberculosis ◽

Antimicrobial Peptides ◽

Experimental Models ◽

New Drugs ◽

World Health ◽

Public Health Issue ◽

Health Organization ◽

Drug Resistant Strains

Tuberculosis is an ancient disease that has become a serious public health issue in recent years, although increasing incidence has been controlled, deaths caused by Mycobacterium tuberculosis have been accentuated due to the emerging of multi-drug resistant strains and the comorbidity with diabetes mellitus and HIV. This situation is threatening the goals of World Health Organization (WHO) to eradicate tuberculosis in 2035. WHO has called for the creation of new drugs as an alternative for the treatment of pulmonary tuberculosis, among the plausible molecules that can be used are the Antimicrobial Peptides (AMPs). These peptides have demonstrated remarkable efficacy to kill mycobacteria in vitro and in vivo in experimental models, nevertheless, these peptides not only have antimicrobial activity but also have a wide variety of functions such as angiogenesis, wound healing, immunomodulation and other well-described roles into the human physiology. Therapeutic strategies for tuberculosis using AMPs must be well thought prior to their clinical use; evaluating comorbidities, family history and risk factors to other diseases, since the wide function of AMPs, they could lead to collateral undesirable effects.

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Phytosomal Curcumin Elicits Anti-tumor Properties Through Suppression of Angiogenesis, Cell Proliferation and Induction of Oxidative Stress in Colorectal Cancer

Current Pharmaceutical Design ◽

10.2174/1381612825666190110145151 ◽

2019 ◽

Vol 24 (39) ◽

pp. 4626-4638 ◽

Cited By ~ 13

Author(s):

Reyhaneh Moradi-Marjaneh ◽

Seyed M. Hassanian ◽

Farzad Rahmani ◽

Seyed H. Aghaee-Bakhtiari ◽

Amir Avan ◽

...

Keyword(s):

Oxidative Stress ◽

Colorectal Cancer ◽

Therapeutic Potential ◽

Vegf Signaling ◽

Anticancer Effects ◽

Inhibition Of Angiogenesis ◽

Underlying Mechanisms ◽

Apoptotic Activity

Background: Colorectal cancer (CRC) is one of the most common causes of cancer-associated mortality in the world. Anti-tumor effect of curcumin has been shown in different cancers; however, the therapeutic potential of novel phytosomal curcumin, as well as the underlying molecular mechanism in CRC, has not yet been explored. Methods: The anti-proliferative, anti-migratory and apoptotic activity of phytosomal curcumin in CT26 cells was assessed by MTT assay, wound healing assay and Flow cytometry, respectively. Phytosomal curcumin was also tested for its in-vivo activity in a xenograft mouse model of CRC. In addition, oxidant/antioxidant activity was examined by DCFH-DA assay in vitro, measurement of malondialdehyde (MDA), Thiol and superoxidedismutase (SOD) and catalase (CAT) activity and also evaluation of expression levels of Nrf2 and GCLM by qRT-PCR in tumor tissues. In addition, the effect of phytosomal curcumin on angiogenesis was assessed by the measurement of VEGF-A and VEGFR-1 and VEGF signaling regulatory microRNAs (miRNAs) in tumor tissue. Results: Phytosomal curcumin exerts anti-proliferative, anti-migratory and apoptotic activity in-vitro. It also decreases tumor growth and augmented 5-fluorouracil (5-FU) anti-tumor effect in-vivo. In addition, our data showed that induction of oxidative stress and inhibition of angiogenesis through modulation of VEGF signaling regulatory miRNAs might be underlying mechanisms by which phytosomal curcumin exerted its antitumor effect. Conclusion: Our data confirmed this notion that phytosomal curcumin administrates anticancer effects and can be used as a complementary treatment in clinical settings.

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