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APOBEC3B expression in adrenocortical carcinoma. (A) TaqMan quantitative real-time PCR of APOBEC3B in
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Recent evidence has implicated APOBEC3B (Apolipoprotein B mRNA editing enzyme catalytic subunit 3B) as a source of mutations in breast, bladder, cervical, lung, head, and neck cancers. However, the role of APOBEC3B in adrenocortical carcinoma (ACC) and the mechanisms through which its expression is regulated in cancer are not fully understood. Here...
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... analyzed APOBEC3B gene expression in 21 normal adrenal cortices, 69 benign adrenocortical tumors and 38 primary ACC samples and found that APOBEC3B mRNA is significantly overexpressed in ACC (p < 0.001) ( Figure 1A). We also analyzed two publicly available datasets from GEO (GSE10927 deposited by Giordano et al.) and EMBL-EBI (E-TABM-311 deposited by Reynies et al.) for the expression of APOBEC3B and observed that in both cohorts, there was a 5-6 fold higher expression in ACC compared to normal and benign adrenocortical tissue samples (p < 0.01) with no difference in expression between benign tumors and normal adrenocortical tissue samples ( Figure 1B and 1C). ...
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... analyzed APOBEC3B gene expression in 21 normal adrenal cortices, 69 benign adrenocortical tumors and 38 primary ACC samples and found that APOBEC3B mRNA is significantly overexpressed in ACC (p < 0.001) ( Figure 1A). We also analyzed two publicly available datasets from GEO (GSE10927 deposited by Giordano et al.) and EMBL-EBI (E-TABM-311 deposited by Reynies et al.) for the expression of APOBEC3B and observed that in both cohorts, there was a 5-6 fold higher expression in ACC compared to normal and benign adrenocortical tissue samples (p < 0.01) with no difference in expression between benign tumors and normal adrenocortical tissue samples ( Figure 1B and 1C). APOBEC3B protein was also overexpressed, by immunofluorescence, in ACC while little or no expression was detected in normal adrenal cortex and benign adrenocortical tumors ( Figure 1D). ...
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... also analyzed two publicly available datasets from GEO (GSE10927 deposited by Giordano et al.) and EMBL-EBI (E-TABM-311 deposited by Reynies et al.) for the expression of APOBEC3B and observed that in both cohorts, there was a 5-6 fold higher expression in ACC compared to normal and benign adrenocortical tissue samples (p < 0.01) with no difference in expression between benign tumors and normal adrenocortical tissue samples ( Figure 1B and 1C). APOBEC3B protein was also overexpressed, by immunofluorescence, in ACC while little or no expression was detected in normal adrenal cortex and benign adrenocortical tumors ( Figure 1D). We evaluated APOBEC3B mRNA and protein expression in three adrenocortical cell lines (BD140, H295R, and SW13) and HEK293 cells, we detected two isoforms of APOBEC3B with a predicted molecular weight of ~43 and ~46 kD ( Figure 1E and 1F). ...
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... protein was also overexpressed, by immunofluorescence, in ACC while little or no expression was detected in normal adrenal cortex and benign adrenocortical tumors ( Figure 1D). We evaluated APOBEC3B mRNA and protein expression in three adrenocortical cell lines (BD140, H295R, and SW13) and HEK293 cells, we detected two isoforms of APOBEC3B with a predicted molecular weight of ~43 and ~46 kD ( Figure 1E and 1F). The RT-qPCR and Western blot analysis showed the highest level of APOBEC3B expression in the H295R cell line as compared to BD140A, SW13, and HEK293 cell lines. ...
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... RT-qPCR and Western blot analysis showed the highest level of APOBEC3B expression in the H295R cell line as compared to BD140A, SW13, and HEK293 cell lines. (Figure 1E and 1F, and Supplementary Figure 1). For subsequent experiements, we used H295R and BD140A cells as they are more representative of ACCs. ...
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... RT-qPCR and Western blot analysis showed the highest level of APOBEC3B expression in the H295R cell line as compared to BD140A, SW13, and HEK293 cell lines. (Figure 1E and 1F, and Supplementary Figure 1). For subsequent experiements, we used H295R and BD140A cells as they are more representative of ACCs. ...
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... gene expression data in Figure 1A-1C, we have performed a one way ANOVA with Sidak's multiple comparisons test. For gene knockdown and cell proliferation experiments in Figure 2A-2D, we have used two way ANOVA statistical testing with Dunnett's correction for multiple comparison. ...
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... analyzed APOBEC3B gene expression in 21 normal adrenal cortices, 69 benign adrenocortical tumors and 38 primary ACC samples and found that APOBEC3B mRNA is significantly overexpressed in ACC (p < 0.001) ( Figure 1A). We also analyzed two publicly available datasets from GEO (GSE10927 deposited by Giordano et al.) and EMBL-EBI (E-TABM-311 deposited by Reynies et al.) for the expression of APOBEC3B and observed that in both cohorts, there was a 5-6 fold higher expression in ACC compared to normal and benign adrenocortical tissue samples (p < 0.01) with no difference in expression between benign tumors and normal adrenocortical tissue samples ( Figure 1B and 1C). ...
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... analyzed APOBEC3B gene expression in 21 normal adrenal cortices, 69 benign adrenocortical tumors and 38 primary ACC samples and found that APOBEC3B mRNA is significantly overexpressed in ACC (p < 0.001) ( Figure 1A). We also analyzed two publicly available datasets from GEO (GSE10927 deposited by Giordano et al.) and EMBL-EBI (E-TABM-311 deposited by Reynies et al.) for the expression of APOBEC3B and observed that in both cohorts, there was a 5-6 fold higher expression in ACC compared to normal and benign adrenocortical tissue samples (p < 0.01) with no difference in expression between benign tumors and normal adrenocortical tissue samples ( Figure 1B and 1C). APOBEC3B protein was also overexpressed, by immunofluorescence, in ACC while little or no expression was detected in normal adrenal cortex and benign adrenocortical tumors ( Figure 1D). ...
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... also analyzed two publicly available datasets from GEO (GSE10927 deposited by Giordano et al.) and EMBL-EBI (E-TABM-311 deposited by Reynies et al.) for the expression of APOBEC3B and observed that in both cohorts, there was a 5-6 fold higher expression in ACC compared to normal and benign adrenocortical tissue samples (p < 0.01) with no difference in expression between benign tumors and normal adrenocortical tissue samples ( Figure 1B and 1C). APOBEC3B protein was also overexpressed, by immunofluorescence, in ACC while little or no expression was detected in normal adrenal cortex and benign adrenocortical tumors ( Figure 1D). We evaluated APOBEC3B mRNA and protein expression in three adrenocortical cell lines (BD140, H295R, and SW13) and HEK293 cells, we detected two isoforms of APOBEC3B with a predicted molecular weight of ~43 and ~46 kD ( Figure 1E and 1F). ...
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... protein was also overexpressed, by immunofluorescence, in ACC while little or no expression was detected in normal adrenal cortex and benign adrenocortical tumors ( Figure 1D). We evaluated APOBEC3B mRNA and protein expression in three adrenocortical cell lines (BD140, H295R, and SW13) and HEK293 cells, we detected two isoforms of APOBEC3B with a predicted molecular weight of ~43 and ~46 kD ( Figure 1E and 1F). The RT-qPCR and Western blot analysis showed the highest level of APOBEC3B expression in the H295R cell line as compared to BD140A, SW13, and HEK293 cell lines. ...
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... RT-qPCR and Western blot analysis showed the highest level of APOBEC3B expression in the H295R cell line as compared to BD140A, SW13, and HEK293 cell lines. (Figure 1E and 1F, and Supplementary Figure 1). For subsequent experiements, we used H295R and BD140A cells as they are more representative of ACCs. ...
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... RT-qPCR and Western blot analysis showed the highest level of APOBEC3B expression in the H295R cell line as compared to BD140A, SW13, and HEK293 cell lines. (Figure 1E and 1F, and Supplementary Figure 1). For subsequent experiements, we used H295R and BD140A cells as they are more representative of ACCs. ...
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Context
Adrenocortical carcinoma (ACC) is a rare aggressive disease with heterogeneous prognosis. Previous studies identified hypermethylation in the promoter region of specific genes to be associated with poor clinical outcome.
Objective
Comparative analysis of promising hypermethylated genes as prognostic markers and evaluation of their added va...
Citations
... 17 APOBEC3B and APOBEC3 signature mutations have also been shown to be upregulated in human cancers. [18][19][20][21] We have previously shown that expression of human APOBEC3B drives resistance to several tumor immunotherapies. One such Molecular Therapy: Oncology therapy that APOBEC3B mediates escape from and resistance to is oncolytic vesicular stomatitis virus (VSV) therapy, 22 which is mediated by a mutation in the RNA-binding cold-shock domain containing E1 protein, a regulator of RNA translation. ...
Chimeric antigen receptor (CAR) T cells are a clinically approved therapy for blood cancers. To produce clinical-grade CAR T cells, a retroviral or lentiviral vector is used to deliver the CAR and associated genes to patient T cells. Apolipoprotein B editing enzyme, catalytic polypeptide 3 (APOBEC3) enzymes are known to be upregulated after transfection and retroviral infection and to deaminate cytidine to uracil in nucleic acids, resulting in cytidine-to-thymine mutations in DNA. Here, we hypothesized that APOBEC3 enzymes, induced during the production of CAR T cells, impact the efficacy of the resulting CAR T cells. We demonstrated that APOBEC3 family member APOBEC3B was upregulated at the RNA and protein levels after transfection of HEK293T cells with plasmids to make lentivirus, and that APOBEC3 signature mutations were present in the CAR construct. APOBEC3B overexpression in HEK293T cells led to further mutations in the resulting CAR T cells, and significantly decreased CAR T cell killing. APOBEC3B knockout in HEK293T cells led to reduced mutations in the CAR construct and significantly increased in CAR T cell killing. These results suggest that generation of CAR-expressing viruses from producer cell lines deficient in genome-modifying proteins such as APOBEC3B could enhance the quality of CAR T cell production.
... David et al. [34] reported that APOBEC3B expression was correlated with adverse clinicopathological features, as well as proliferative characteristics, such as cell cycle-related genes and mitosis, in breast cancer. Du et al. [35] reported that APOBEC3B knockdown affected ovarian cancer cell viability and, in adrenal cortical cancer [36], knockdown of APOBEC3B reduced cell proliferation and tumor growth in a xenograft mouse model. However, the mechanism by which APOBEC3B affects GC progression remains elusive. ...
... The mechanisms by which APOBEC3B is dysregulated in multiple cancers remain unclear. Gara et al. [36] reported that high APOBEC3B expression results in the increase/loss of chromosomes 4 and 8, as well as an increase in P53 mutation frequency, in adrenal cortical carcinoma. P53 is critically involved in APOBEC3B regulation. ...
Gastric cancer is a common malignant tumor with a high mortality rate. Abnormal APOBEC3B (apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3B) expression increases tumor susceptibility. However, the exact molecular mechanism of APOBEC3B expression in the development of gastric cancer is still unknown. We investigated the effect of APOBEC3B on the malignant biological behavior of gastric cancer cells and discussed the role of APOBEC3B in the development and progression of gastric cancer. APOBEC3B protein levels were measured in 161 gastric cancer samples using western blotting and immunohistochemistry. Both in vitro and in vivo assays were performed, and molecules were analyzed using bioinformatics analysis and western blotting. APOBEC3B was overexpressed in gastric cancer. Moreover, APOBEC3B significantly enhanced cell proliferation in vitro and tumorigenicity in vivo. Regarding the underlying mechanism, APOBEC3B promoted the proliferation of gastric cancer cells by upregulating P53, MCM2 (minichromosome maintenance protein 2), and cyclin D1. Our results suggest that APOBEC3B is involved in cancer progression, providing a new theoretical basis for the prevention and treatment of gastric cancer.
... Accordingly, TP53 mutations-which are mostly non-APOBEC3-induced-were more common in bladder cancers, lung adenocarcinomas, and B-cell lymphoma cell lines with high burden of APOBEC3-induced mutations [46,89,149]. Similarly, high APOBEC3B expression has been associated with more p53 mutations in breast cancer and adrenocortical carcinoma [22,44]. In addition to selection pressure, this trend could arise from higher APOBEC3 expression in p53-mutated tumors since p53 may suppress APOBEC3B transcription via p21 and DREAM proteins [150]. ...
... Immunosuppression may synergize with APOBEC3mediated mutagenesis in some cancers, allowing oncogenic mutations to accumulate and drive tumorigenesis while averting the host immune response. In accordance with this model, studies in adrenocortical carcinoma and gastric cancer-which both show immunosuppression with APOBEC3s-have found that higher APOBEC3B expression is correlated with shorter survival (Tables 4 and 5) [37,38,44]. Elevated APOBEC3 expression has also been associated with adverse clinical outcomes in nasopharyngeal carcinoma, clear cell renal carcinoma, and neuroendocrine tumors, though little is known about APOBEC3-mediated immune effects in these cancers (Table 5) [55,169]. ...
Apolipoprotein B mRNA-editing enzyme, catalytic polypeptides (APOBECs) are cytosine deaminases involved in innate and adaptive immunity. However, some APOBEC family members can also deaminate host genomes to generate oncogenic mutations. The resulting mutations, primarily signatures 2 and 13, occur in many tumor types and are among the most common mutational signatures in cancer. This review summarizes the current evidence implicating APOBEC3s as major mutators and outlines the exogenous and endogenous triggers of APOBEC3 expression and mutational activity. The review also discusses how APOBEC3-mediated mutagenesis impacts tumor evolution through both mutagenic and non-mutagenic pathways, including by inducing driver mutations and modulating the tumor immune microenvironment. Moving from molecular biology to clinical outcomes, the review concludes by summarizing the divergent prognostic significance of APOBEC3s across cancer types and their therapeutic potential in the current and future clinical landscapes.
... Although deregulation of APOBEC3B has been reported in many cancer types, the conclusions about the prognostic value of A3B for patients remain controversial. A large number of studies reported that high A3B expression is linked with adverse outcomes or poor prognosis in cancer patients [40,[42][43][44]. However, several studies reported that A3B expression improved OS in patients with cancer or have no correlations with disease-free or overall survival [32,45,46]. ...
Introduction:
Controversy exists regarding the association of apolipoprotein B mRNA editing enzyme catalytic subunit 3B APOBEC3B, (A3B) overexpression and poor prognosis, metastasis, and chemotherapy drug resistance in cancers. Here we conducted a systematic review and meta-analysis to determine its prognostic value and clinicopathological features in breast cancer and some other malignancies.
Materials and methods:
PubMed, Scopus, Cochrane Library, Web of Science, and EMBASE were searched up to Feb 2022 for the association of A3B with breast, ovarian, gastrointestinal and lung cancers. The pooled hazard ratios with 95% confidence interval (CI) were evaluated to assess disease-free survival (DFS), overall survival (OS), and recurrence-free survival (RFS) in cancers under study.
Results:
Over 3700 patients were included in this meta-survey. Elevated levels of A3B were significantly related to low OS (pooled HR = 1.30; 95% CI:1.09-1.55, P < 0.01), poor DFS (pooled HR = 1.66; 95% CI:1.17-2.35, P < 0.01) and poor RFS (HR = 1.51, 95% CI:1.11-2.04, P = 0.01). Subgroup analysis revealed that high A3B expression was associated with poor OS in lung (HR = 1.85, 95% CI: 1.40-2.45), and breast cancers (HR = 1.38, 95% CI: 1.00-1.89). High expression of A3B did not display any significant association with clinicopathologic features.
Conclusion:
APOBEC3B overexpression is related to poor OS, DFS and RFS only in some cancer types and no generalized role could be predicted for all cancers.
... Thus, in the case of multiple myeloma, a high level of APOBECmediated transitions gives an unfavorable prognosis [113,114]. A similar observation was made for APO-BEC3B in cohorts of patients with adrenal carcinoma selected from the Cancer Genome Atlas database [115] and patients with ovarian cancer [116]. ...
Proteins of the AID/APOBEC family are capable of cytidine deamination in nucleic acids forming uracil. These enzymes are involved in mRNA editing, protection against viruses, the introduction of point mutations into DNA during somatic hypermutation, and antibody isotype switching. Since these deaminases, especially AID, are potent mutagens, their expression, activity, and specificity are regulated by several intracellular mechanisms. In this review, we discuss the mechanisms of impaired expression and activation of AID/APOBEC proteins in human tumors and their role in carcinogenesis and tumor progression. Also, the diagnostic and potential therapeutic value of increased expression of AID/APOBEC in different types of tumors is analyzed. We assume that in the case of solid tumors, increased expression of endogenous deaminases can serve as a marker of response to immunotherapy since multiple point mutations in host DNA could lead to amino acid substitutions in tumor proteins and thereby increase the frequency of neoepitopes.
... For example, significantly higher risk and consequent reduced OS and DFS were detected for KIRP patients expressing MEOX1 and MEOX2 (Additional file 3: Fig. S2) [53]. A similar result was also found for adrenocortical adenocarcinoma (ACC) patients expressing GATA3, GATA2, and SOX11 (Additional file 4: Fig. S3) [54]. ...
Aim:
Neuroblastoma is a heterogeneous childhood cancer derived from the neural crest. The dual cell identities of neuroblastoma include Mesenchymal (MES) and Adrenergic (ADRN). These identities are conferred by a small set of tightly-regulated transcription factors (TFs) binding super enhancers, collectively forming core regulatory circuitries (CRCs). The purpose of this study was to gain a deep understanding of the role of MES and ADRN TFs in neuroblastoma and other cancers as potential indicators of disease prognosis, progression, and relapse.
Methods:
To that end, we first investigated the expression and mutational profile of MES and ADRN TFs in neuroblastoma. Moreover, we established their correlation with neuroblastoma risk groups and overall survival while establishing their extended networks with long non-coding RNAs (lncRNAs). Furthermore, we analysed the pan-cancer expression and mutational profile of these TFs and their correlation with patient survival and finally their network connectivity, using a panel of bioinformatic tools including GEPIA2, human pathology atlas, TIMER2, Omicsnet, and Cytoscape.
Results:
We show the association of multiple MES and ADRN TFs with neuroblastoma risk groups and overall survival and find significantly higher expression of various MES and ADRN TFs compared to normal tissues and their association with overall survival and disease-free survival in multiple cancers. Moreover, we report the strong correlation of the expression of these TFs with the infiltration of stromal and immune cells in the tumour microenvironment and with stemness and metastasis-related genes. Furthermore, we reveal extended pan-cancer networks comprising these TFs that influence the tumour microenvironment and metastasis and may be useful indicators of cancer prognosis and patient survival.
Conclusion:
Our meta-analysis shows the significance of MES and ADRN TFs as indicators of patient prognosis and the putative utility of these TFs as potential novel biomarkers.
... Recent research has also shown that the expression levels of A3B have a strong positive correlation with tumor mutation loads (11), along with a strong A3B upregulation in the majority of tumor types, including breast, uterus, bladder, head and neck, and lung (adeno-and squamous cell carcinomas) (12), suggesting that A3B may be a general endogenous mutagen that contributes to human cancers. Patients with a higher level of A3B gene expression in their tumors had a lower survival rate as compared to patients with lower expression A3B gene expression in their tumors (13)(14)(15). ...
Background:
Approximately 80-85% of lung cancer is the non-small cell lung cancer (NSCLC) subtype, which ranks as the leading cause of cancer deaths worldwide. APOBEC3B (A3B) was reported to be a key source of mutations in NSCLC. However, the role of the A3B deletion polymorphism in the etiology of NSCLC has not been well-documented.
Methods:
A case-control study with 317 NSCLC patients and 334 healthy controls was conducted to explore the association between the A3B deletion polymorphism and the risk of NSCLC. The unconditional logistic regression model was performed to calculate the odds ratio (OR) and the 95% confidence interval (CI), and the confounding factors were adjusted, including age, gender, and smoking status, to estimate the risk. An analysis of gene-environment interactions was performed using multifactor dimensionality reduction (MDR) software.
Results:
We found that the del/del genotype of A3B deletion significantly increased NSCLC risk. Compared with individuals carrying the ins/ins genotype of A3B deletion, individuals with the del/del genotype had a 2.36 times increased risk of developing NSCLC after adjusting for confounding factors (OR =2.71, 95% CI: 1.67-4.42, P<0.001). A 3-factor gene-environment (A3B deletion, gender, and smoking) interaction model was found for NSCLC (OR =4.407, 95% CI: 1.174-16.549, P=0.028).
Conclusions:
We propose that the A3B deletion polymorphism can increase the risk of developing NSCLC, and their interactions with gender and smoking may contribute to the risk of NSCLC in the southern Chinese population.
... Next generation DNA sequencing (NGS) has brought about revolution in genomic research. In the literature, numerous reports can be found proposing comprehensive and novel molecular pathophysiological, diagnostic, therapeutic and prognostic genes associated with ACC [4,50,113,[141][142][143][144][145][146][147][148][149][150][151][152][153][154][155][156][157][158][159][160]. Zheng et al. have summarized proposed genes as potential drivers involved in sporadic adrenocortical tumorigenesis, including insulin-like growth factor 2, β-catenin (CTNNB1), TP53, ZNRF3 and TERT, as well as novel nominated drivers such as PRKAR1A, RPL22, TERF2, CCNE1 and NF1 [161]. ...
Adrenocortical carcinoma (ACC) is a rare endocrine malignancy arising from the adrenal cortex often with unexpected biological behavior. It can occur at any age, with two peaks of incidence: in the first and between fifth and seventh decades of life. Although ACC are mostly hormonally active, precursors and metabolites, rather than end products of steroidogenesis are produced by dedifferentiated and immature malignant cells. Distinguishing the etiology of adrenal mass, between benign adenomas, which are quite frequent in general population, and malignant carcinomas with dismal prognosis is often unfeasible. Even after pathohistological analysis, diagnosis of adrenocortical carcinomas is not always straightforward and represents a great challenge for experienced and multidisciplinary expert teams. No single imaging method, hormonal work-up or immunohistochemical labelling can definitively prove the diagnosis of ACC. Over several decades’ great efforts have been made in finding novel reliable and available diagnostic and prognostic factors including steroid metabolome profiling or target gene identification. Despite these achievements, the 5-year mortality rate still accounts for approximately 75% to 90%, ACC is frequently diagnosed in advanced stages and therapeutic options are unfortunately limited. Therefore, imperative is to identify new biological markers that can predict patient prognosis and provide new therapeutic options.
Adrenocortical carcinoma (ACC) is a rare endocrine malignancy with a poor prognosis and limited treatment options. Bulk genomic characterization of ACC has not yielded obvious therapeutic or immunotherapeutic targets, yet novel therapies are needed. We hypothesized that elucidating the intratumoral cellular heterogeneity by single nuclei RNA sequencing analyses would yield insights into potential therapeutic vulnerabilities of this disease. In addition to characterizing the immune cell and fibroblast landscape, our analyses of single nuclei gene expression profiles identified an adrenal cortex cell cluster exhibiting a program of replication stress and DNA damage response in primary and metastatic ACC. In vitro assessment of replication stress and DNA damage response using an ACC cell line and a series of newly-derived hormonally active patient-derived tumor organoids revealed ATR sensitivity. These findings provide novel mechanistic insight into ACC biology and suggest that an underlying dependency on ATR may be leveraged therapeutically in advanced ACC.
