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BMJ Open Gastroenterology Aug 2020Differentiating between malignant and normal cells within tissue samples is vital for molecular profiling of cancer using advances in genomics and transcriptomics.... (Comparative Study)
Comparative Study
BACKGROUND
Differentiating between malignant and normal cells within tissue samples is vital for molecular profiling of cancer using advances in genomics and transcriptomics. Cell-surface markers of tumour-normal discrimination have additional value in terms of translatability to diagnostic and therapeutic strategies. In gastric cancer (GC), previous studies have identified individual genes or proteins that are upregulated in cancer. However, a systematic analysis of cell-surface markers and development of a composite panel involving multiple candidates to differentiate tumour from normal has not been previously reported.
METHODS
Whole transcriptome sequencing (WTS) of GC and matched normal samples from the Singapore Gastric Cancer Consortium (SGCC) was used as a discovery cohort to identify upregulated putative cell-surface proteins. Matched WTS data from the The Cancer Genome Atlas (TCGA) was used as a validation cohort. Promising candidates from this analysis were validated orthogonally using multispectral immunohistochemistry (mIHC) with automated quantitative analysis using the Vectra platform. mIHC was performed on a tissue microarray containing matched normal, marginal and tumour tissues. The receiver-operating characteristic (ROC) curves were analysed to identify markers with the highest diagnostic validity independently and in combination.
RESULTS
Analysis of putative membrane protein transcripts from the SGCC discovery cohort WTS data (n=15 matched tumour and normal pairs) identified several differentially and highly expressed candidates in tumour compared with normal tissues. After validation with TCGA data (n=29 matched tumour and normal pairs), the following proteins were selected for mIHC analysis: CEACAM5, CEACAM6, CLDN4, CLDN7, and EpCAM. These were compared with established glycoprotein markers in GC, namely CA19-9 and CA72-4. Individual ROC curves yielded the best performance for CEACAM5 (area under the ROC curve (AUC)=0.80), CEACAM6 (AUC=0.82), EpCAM (AUC=0.83), and CA72-4 (AUC=0.76). Combined multiplexed imaging of these four markers revealed improved specificity and sensitivity for detection of tumour from normal tissue (AUC of 4-plex=0.91).
CONCLUSION
CEAMCAM5, CEACAM6, EpCAM, and CA72-4 form a versatile set of markers for robust discrimination of GC from adjacent normal tissue. As cell-surface markers, they are compatible with both IHC and live imaging approaches. These candidates may be exploited to improve automated identification of tumour tissue in GC.
Topics: Adenocarcinoma; Antigens, CD; Antigens, Tumor-Associated, Carbohydrate; CA-19-9 Antigen; Carcinoembryonic Antigen; Cell Adhesion Molecules; Claudin-4; Claudins; Epithelial Cell Adhesion Molecule; Evaluation Studies as Topic; GPI-Linked Proteins; Genomics; Humans; Immunohistochemistry; Membrane Proteins; ROC Curve; Sensitivity and Specificity; Singapore; Stomach Neoplasms; Up-Regulation; Exome Sequencing
PubMed: 32816956
DOI: 10.1136/bmjgast-2020-000452 -
Iranian Journal of Allergy, Asthma, and... Feb 2022Program cell death protein 1 (PD1) is considered as an inhibitory molecule that is expressed on the surface of activated T-cells and bound to PD-L1 and PD-L2 ligands.... (Review)
Review
Program cell death protein 1 (PD1) is considered as an inhibitory molecule that is expressed on the surface of activated T-cells and bound to PD-L1 and PD-L2 ligands. Several types of cancer cells express PD-L1 which can bind to PD1 on the surface of tumor-specific T-cells. PD1/PD-L1 ligation triggers a pathway to protect tumor cells from an effective response of tumor-specific T-cells. Different PD1/PD-L1 blocker antibodies are clinically used to promote the T-cell response against the cancer cells. Current studies suggest that the gut microbiome impacts the efficiency of PD1 blockade therapy in cancer patients. The association of several bacterial species with PD1 responder patients has been determined. The present study reviewed previous reports on the relation between the microbiome and immune checkpoint therapy (ICT). The results of studies were discussed considering adjuvant and molecular mimicry of microbial antigens by tumor-associated antigens and metabolic effects of microbial products on ICT.
Topics: B7-H1 Antigen; Gastrointestinal Microbiome; Humans; Immunotherapy; Neoplasms; Programmed Cell Death 1 Ligand 2 Protein; Programmed Cell Death 1 Receptor
PubMed: 35524371
DOI: 10.18502/ijaai.v21i1.8607 -
BMC Cancer Sep 2022Trophoblast cell-surface antigen 2 (TROP2) is related to tumor proliferation enhancement and poor prognosis. An antibody targeting TROP2 was developed to treat...
BACKGROUND
Trophoblast cell-surface antigen 2 (TROP2) is related to tumor proliferation enhancement and poor prognosis. An antibody targeting TROP2 was developed to treat metastatic triple-negative breast cancer (TNBC) which has a limited treatment modality. To characterize the TROP2 expressing tumors in TNBC, we analyzed TROP2 expression in three cohorts; (1) primary tumor without neoadjuvant chemotherapy, (2) primary tumor with neoadjuvant chemotherapy, and (3) metastatic tumor.
METHODS
A total of 807 TNBC cases were evaluated for TROP2 immunohistochemical expression. We evaluated the TROP2 H-score distribution in the three cohorts. Tumors were divided into two groups based on TROP2 expression (high vs. low). We analyzed the relationship between clinicopathologic features and markers, including epidermal growth factor receptor, cytokeratin 5/6, p53, and Ki-67, and prognostic significance at high vs. low TROP2 expression.
RESULTS
There was no difference in TROP2 H-score distribution between the three cohorts. Moderate-to-strong membranous expression of TROP2 in at least 10% of tumor cells was present in 662 cases (82.0%) in Cohort 1, 59 cases (89.4%) in Cohort 2, and 23 cases (88.5%) in Cohort 3. There was no significant difference in clinicopathologic features between high vs. low TROP2 in all cohorts. TROP2 H-score was an independent poor prognostic factor for overall survival in Cohort 3.
CONCLUSIONS
TNBC showed similar TROP2 expression regardless of neoadjuvant treatment or primary tumor/metastasis. Although the prognostic significance of TROP2 expression in metastatic TNBC has been revealed, further evaluation of the predictive value of TROP2 expression for targeted therapy is needed.
Topics: Antigens, Neoplasm; Antigens, Surface; Biomarkers, Tumor; Cell Adhesion Molecules; ErbB Receptors; Humans; Keratin-5; Ki-67 Antigen; Prognosis; Triple Negative Breast Neoplasms; Trophoblasts; Tumor Suppressor Protein p53
PubMed: 36153494
DOI: 10.1186/s12885-022-10076-7 -
Journal of Preventive Medicine and... Mar 2022Several diseases are reported to be associated with ABO/Rh blood groups. Data on the association between ABO and Rh D blood group antigens in the Nigerian population is...
BACKGROUND
Several diseases are reported to be associated with ABO/Rh blood groups. Data on the association between ABO and Rh D blood group antigens in the Nigerian population is sparse. This study aimed at determining the prevalence of Hepatitis B Virus (HBV) infection as well as its association with ABO and Rh D antigens among young Nigerian adults.
METHODS
Whole blood was collected from 496 students and screened for the presence of HBsAg using an immuno-chromatographic technique. The ABO and Rh D antigen status of participants were also determined using standard techniques.
RESULTS
In this study, the prevalence of HBV infection was 10/496 (2.10%). Of all factors assessed, only age of participants was identified as a risk factor (P < 0.05) for HBV seropositivity. Over half 257/496 (51.5%) of subjects were of the blood group O type, while 18/496 (3.6%) were of the AB blood type which was the least in occurrence. Rh D negative blood group was observed among 24/496 (4.8%) subjects. Those with the B blood type were observed to have an insignificantly (P > 0.05) higher prevalence of HBV infection. However, with respect to Rh D antigen alone, participants negative for the antigen were observed to have a five times higher risk of acquiring HBV infection than those positive for it (OR = 5.273, 95% CI = 1.056, 26.321, P > 0.05). Combining the ABO and Rh blood group systems, an association (OR = 20.174; P > 0.05) was found to exist between B Rh D negative status and HBV infection.
CONCLUSION
Possession of B antigen without Rh D antigen is associated with increased risk of acquiring HBV infection.
Topics: ABO Blood-Group System; Adult; Hepatitis B; Hepatitis B Surface Antigens; Hepatitis B virus; Humans; Nigeria
PubMed: 35647381
DOI: 10.15167/2421-4248/jpmh2022.63.1.1967 -
PLoS Genetics Jun 2022The Human Leukocyte Antigen (HLA) is a critical genetic system for different outcomes after solid organ and hematopoietic cell transplantation. Its polymorphism is...
The Human Leukocyte Antigen (HLA) is a critical genetic system for different outcomes after solid organ and hematopoietic cell transplantation. Its polymorphism is usually determined by molecular technologies at the DNA level. A potential role of HLA allelic expression remains under investigation in the context of the allogenic immune response between donors and recipients. In this study, we quantified the allelic expression of all three HLA class I loci (HLA-A, B and C) by RNA sequencing and conducted an analysis of expression quantitative traits loci (eQTL) to investigate whether HLA expression regulation could be associated with non-coding gene variations. HLA-B alleles exhibited the highest expression levels followed by HLA-C and HLA-A alleles. The max fold expression variation was observed for HLA-C alleles. The expression of HLA class I loci of distinct individuals demonstrated a coordinated and paired expression of both alleles of the same locus. Expression of conserved HLA-A~B~C haplotypes differed in distinct PBMC's suggesting an individual regulated expression of both HLA class I alleles and haplotypes. Cytokines TNFα /IFNβ, which induced a very similar upregulation of HLA class I RNA and cell surface expression across alleles did not modify the individually coordinated expression at the three HLA class I loci. By identifying cis eQTLs for the HLA class I genes, we show that the non-coding eQTLs explain 29%, 13%, and 31% of the respective HLA-A, B, C expression variance in unstimulated cells, and 9%, 23%, and 50% of the variance in cytokine-stimulated cells. The eQTLs have significantly higher effect sizes in stimulated cells compared to unstimulated cells for HLA-B and HLA-C genes expression. Our data also suggest that the identified eQTLs are independent from the coding variation which defines HLA alleles and thus may be influential on intra-allele expression variability although they might not represent the causal eQTLs.
Topics: Alleles; Gene Frequency; HLA Antigens; HLA-A Antigens; HLA-B Antigens; HLA-C Antigens; Haplotypes; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Humans; Leukocytes, Mononuclear
PubMed: 35666741
DOI: 10.1371/journal.pgen.1010212 -
Cellular and Molecular Gastroenterology... 2023Hepatitis B virus (HBV) was identified as an enveloped DNA virus with a diameter of 42 nm. Multivesicular bodies play a central role in HBV egress and exosome...
BACKGROUND & AIMS
Hepatitis B virus (HBV) was identified as an enveloped DNA virus with a diameter of 42 nm. Multivesicular bodies play a central role in HBV egress and exosome biogenesis. In light of this, it was studied whether intact virions wrapped in exosomes are released by HBV-producing cells.
METHODS
Robust methods for efficient separation of exosomes from virions were established. Exosomes were subjected to limited detergent treatment for release of viral particles. Electron microscopy of immunogold labeled ultrathin sections of purified exosomes was performed for characterization of exosomal HBV. Exosome formation/release was affected by inhibitors or Crispr/Cas-mediated gene silencing. Infectivity/uptake of exosomal HBV was investigated in susceptible and non-susceptible cells.
RESULTS
Exosomes could be isolated from supernatants of HBV-producing cells, which are characterized by the presence of exosomal and HBV markers. These exosomal fractions could be separated from the fractions containing free virions. Limited detergent treatment of exosomes causes stepwise release of intact HBV virions and naked capsids. Inhibition of exosome morphogenesis impairs the release of exosome-wrapped HBV. Electron microscopy confirmed the presence of intact virions in exosomes. Moreover, the presence of large hepatitis B virus surface antigen on the surface of exosomes derived from HBV expressing cells was observed, which conferred exosome-encapsulated HBV initiating infection in susceptible cells in a , large hepatitis B virus surface antigen/Na-taurocholate co-transporting polypeptide-dependent manner. The uptake of exosomal HBV with low efficiency was also observed in non-permissive cells.
CONCLUSION
These data indicate that a fraction of intact HBV virions can be released as exosomes. This reveals a so far not described release pathway for HBV.
Topics: Humans; Exosomes; Detergents; Virion; Hepatitis B; Hepatitis B virus; Antigens, Surface
PubMed: 36184032
DOI: 10.1016/j.jcmgh.2022.09.012 -
Hitting the complexity of the TIGIT-CD96-CD112R-CD226 axis for next-generation cancer immunotherapy.BMB Reports Jan 2021Antibody-based therapeutics targeting the inhibitory receptors PD-1, PD-L1, or CTLA-4 have shown remarkable clinical progress on several cancers. However, most patients... (Review)
Review
Antibody-based therapeutics targeting the inhibitory receptors PD-1, PD-L1, or CTLA-4 have shown remarkable clinical progress on several cancers. However, most patients do not benefit from these therapies. Thus, many efforts are being made to identify new immune checkpoint receptor-ligand pathways that are alternative targets for cancer immunotherapies. Nectin and nectin-like molecules are widely expressed on several types of tumor cells and play regulatory roles in T- and NK-cell functions. TIGIT, CD226, CD96 and CD112R on lymphoid cells are a group of immunoglobulin superfamily receptors that interact with Nectin and nectin-like molecules with different affinities. These receptors transmit activating or inhibitory signals upon binding their cognate ligands to the immune cells. The integrated signals formed by their complex interactions contribute to regulating immune-cell functions. Several clinical trials are currently evaluating the efficacy of anti-TIGIT and anti-CD112R blockades for treating patients with solid tumors. However, many questions still need to be answered in order to fully understand the dynamics and functions of these receptor networks. This review addresses the rationale behind targeting TIGIT, CD226, CD96, and CD112R to regulate T- and NK-cell functions and discusses their potential application in cancer immunotherapy. [BMB Reports 2021; 54(1): 2-11].
Topics: Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Humans; Immunotherapy; Neoplasms; Receptors, Cell Surface; Receptors, Immunologic
PubMed: 33298247
DOI: 10.5483/BMBRep.2021.54.1.229 -
Biology Open Jul 2022Epithelial cell adhesion molecule (EPCAM) is a transmembrane glycoprotein expressed on the surface of most epithelial and epithelium-derived tumor cells and reported to...
Epithelial cell adhesion molecule (EPCAM) is a transmembrane glycoprotein expressed on the surface of most epithelial and epithelium-derived tumor cells and reported to regulate stability of epithelial tight junction proteins, claudins. Despite its widespread expression, loss of EPCAM function has so far only been reported to prominently affect intestinal development, resulting in severe early onset enteropathy associated with impaired growth and decreased survival in both humans and mice. In this study, we show that the critical role of EPCAM is not limited to intestinal tissues and that it shares its essential function with its only known homolog, Trophoblast cell surface antigen 2 (TROP2). EPCAM-deficient mice show significant growth retardation and die within 4 weeks after birth. In addition to changes in small and large intestines, loss of EPCAM results in hyperkeratosis in the skin and forestomach, hair follicle atrophy leading to alopecia, nephron hypoplasia in the kidney, proteinuria, and altered production of digestive enzymes by the pancreas. Expression of TROP2 partially, but not completely, overlaps with EPCAM in a number developing epithelia. Although loss of TROP2 had no gross impact on mouse development and survival, TROP2 deficiency generally compounded developmental defects observed in EPCAM-deficient mice, led to an approximately 60% decrease in embryonic viability, and further shortened postnatal lifespan of born pups. Importantly, TROP2 was able to compensate for the loss of EPCAM in stabilizing claudin-7 expression and cell membrane localization in tissues that co-express both proteins. These findings identify overlapping functions of EPCAM and TROP2 as regulators of epithelial development in both intestinal and extraintestinal tissues.
Topics: Animals; Antigens, Neoplasm; Cell Adhesion Molecules; Claudins; Epithelial Cell Adhesion Molecule; Epithelium; Intestines; Mice
PubMed: 35730316
DOI: 10.1242/bio.059403 -
Hepatitis B virus middle surface antigen loss promotes clinical variant persistence in mouse models.Virulence Dec 2021Hepatitis B virus (HBV) middle surface antigen (MHBs) mutation or deletion occurs in patients with chronic HBV infection. However, the functional role of MHBs in HBV...
Hepatitis B virus (HBV) middle surface antigen (MHBs) mutation or deletion occurs in patients with chronic HBV infection. However, the functional role of MHBs in HBV infection is still an enigma. Here, we reported that 7.33% (11/150) isolates of CHB patients had MHBs start codon mutations compared with 0.00% (0/146) in acute hepatitis B (AHB) patients. Interestingly, MHBs loss accounted for 11.88% (126/1061) isolates from NCBI GenBank, compared with 0.09% (1/1061) and 0.00% (0/1061) for HBV large surface antigen (LHBs) loss and HBV small surface antigen (SHBs) loss, respectively. One persistent HBV clone of genotype B (B56, MHBs loss) from a CHB patient was hydrodynamically injected into BALB/c mice. B56 persisted for >70 weeks in BALB/c mice, whereas B56 with restored MHBs (B56) was quickly cleared within 28 days. Serum cytokine assays demonstrated that CXCL1, CXCL2, IL-6 and IL-33 were significantly increased during rapid HBV clearance in B56 mice. Furthermore, the enhancers and promoters of B56 were proved to be required for B56 persistence in mice. Ablating MHBs expression improved the persistence of a new clone (HBV1.3, genotype B) which was recreated by using enhancers and promoters of B56. These data demonstrated that MHBs deletion can promote the persistence of specific HBV variants in a hydrodynamic mouse model. MHBs re-expression restored a rapid clearance of HBV, which was accompanied by cytokine responses including the elevation of CXCL1, CXCL2, IL-6 and IL-33.
Topics: Animals; Antigens, Surface; Disease Models, Animal; Hepatitis B Surface Antigens; Hepatitis B virus; Humans; Interleukin-33; Interleukin-6; Mice; Mice, Inbred BALB C
PubMed: 34738866
DOI: 10.1080/21505594.2021.1999130 -
Nature Communications May 2022Hepatitis B virus has infected a third of the world's population, and 296 million people are living with chronic infection. Chronic infection leads to progressive liver...
Hepatitis B virus has infected a third of the world's population, and 296 million people are living with chronic infection. Chronic infection leads to progressive liver disease, including hepatocellular carcinoma and liver failure, and there remains no reliable curative therapy. These gaps in our understanding are due, in large part, to a paucity of animal models of HBV infection. Here, we show that rhesus macaques regularly clear acute HBV infection, similar to adult humans, but can develop long-term infection if immunosuppressed. Similar to patients, we longitudinally detected HBV DNA, HBV surface antigen, and HBV e antigen in the serum of experimentally infected animals. In addition, we discovered hallmarks of HBV infection in the liver, including RNA transcription, HBV core and HBV surface antigen translation, and covalently closed circular DNA biogenesis. This pre-clinical animal model will serve to accelerate emerging HBV curative therapies into the clinic.
Topics: Animals; Antigens, Surface; Hepatitis B; Hepatitis B virus; Hepatitis B, Chronic; Humans; Liver Neoplasms; Macaca mulatta
PubMed: 35637225
DOI: 10.1038/s41467-022-30593-0