Diagnostic and prognostic potential of the intra-tumoral microbiota profile in HPV-independent endocervical adenocarcinoma

BackgroundMicrobial community dynamics have been involved in numerous diseases, including cancer. The diversity of intertumoral microbiota in human papillomavirus independent endocervical adenocarcinoma (HPVI ECA) is not well-characterized.ObjectiveOur objective is to delineate the intratumoral microbiota profile in HPVI ECA and investigate its potential influence on oncogenesis.MethodsWe analyzed 45 HPVI ECA cases, comprising 36 gastric-type ECA (GEA) and 9 clear cell carcinomas (CCC). We compared the microbial composition within cancerous and adjacent noncancerous tissue samples using 5R-16S ribosomal DNA sequencing. Further, we investigated the correlation between specific microbes and clinical-pathological metrics as well as patient outcomes.ResultsOur findings demonstrate notable differences in the microbial spectra between cancerous and adjacent noncancerous tissues. Amongst HPVI ECA subtypes, GEAs exhibit more microbial variations compared to CCCs. Using the Random Forest algorithm, we identified two distinct microbial signatures that could act as predictive biomarkers for HPVI ECA and differentiate between GEA and CCC. Varied microbial abundances was related to clinical characteristics of HPVI ECA patients. In addition, high levels of Micrococcus and low levels of unknown genus75 from the Comamonadaceae family were associated with poorer outcomes in HPVI ECA patients. Similarly, an abundance of Microbacterium correlated with reduced overall survival (OS), and a high presence of Streptococcaceae family microbes was linked to reduced recurrence-free survival (RFS) in GEA patients. Intriguingly, a high abundance of Micrococcus was also associated with a worse OS in GEA patients.ConclusionThe study reveals distinct microbial signatures in HPVI ECA, which have potential as biomarkers for disease prognosis. The correlation between these tumor-associated microbiota features and clinicopathological characteristics underscores the possibility of microbiome-based interventions. Our research provides a foundation for more in-depth studies into the cervical microbiome’s role in HPVI ECA

Abstract B38: Cancer-related circulating long noncoding RNAs in serous extracellular vesicles: Their characterization and potential application

Abstract Background: Circulating nucleic acids (CNAs) are extracellular nucleic acids found in cell-free sera, plasma and other bodily fluids of healthy subjects and cancer patients. We have previously demonstrated that human serum or plasma contains microRNAs (miRNAs) and long non-coding RNA (lncRNAs) that are significantly up-regulated or down-regulated in various types of cancer and are of good diagnostic value for screening. The mechanisms underlying the stability of serum RNAs are unclear; lncRNAs may be protected by extracellular vesicles (EV) including apoptotic body (AB), microvesicle (MV) and exosome (EXO), as for circulating miRNAs. In order to find optimal method to evaluate the potential utility of circulating lncRNAs for cancer diagnosis or prognosis, more exploration should be undertook for the distribution of circulating lncRNAs. Results: The shapes and sizes of three subgroups of extracellular vesicles, including AB, MV and EXO, were evaluated. NanoSight particle tracking analysis and transmission electron microscopy (TEM) showed the MVs, pelleted at 12, 000×g, were with the size range of 75-465 nm; and EXOs, pelleted at 120,000×g, with the size range of 45-205 nm. In serous vesicles of both colorectal cancer and healthy subjects, 17 of the 39 cancer-related lncRNAs were detected, including TDRG1, MEG3, DSCAM-AS1, 7SK, MIR31HG, CBR3-AS1, TUG1, BCAR4, EPB41L4A, PCA3, FAS-AS1, SUMO1P3, uc338, PRNCR1, PTENP1 and CUDR. The amount of most lncRNAs were higher in EXO than those in AB and MV. In addition, AB contains higher amount of most lncRNAs than MV. Results: The shapes and sizes of three subgroups of extracellular vesicles, including AB, MV and EXO, were evaluated. NanoSight particle tracking analysis and transmission electron microscopy (TEM) showed the MVs, pelleted at 12, 000×g, were with the size range of 75-465 nm; and EXOs, pelleted at 120,000×g, with the size range of 45-205 nm. In sera of both colorectal cancer and healthy subjects, 16 out of the 39 cancer-related lncRNAs were detected. The amount of most lncRNAs were higher in EXO than those in AB and MV. In addition, AB contains higher amount of most lncRNAs than MV. Conclusions: Among three types of vesicles in sera, EXOs were the richest reservoir for almost all measured lncRNAs, while the MVs contained the least amount of lncRNAs. EXOs, as their intercellular origin, have reasons to contain higher amount of lncRNAs among serous vesicles and seem to be the most promising research materials in the field of circulating lncRNA. Citation Format: Dong Lei, Lin Wanrun, Qi Peng, Xu Midie, Du Xiang. Cancer-related circulating long noncoding RNAs in serous extracellular vesicles: Their characterization and potential application. [abstract]. In: Proceedings of the AACR Special Conference on Noncoding RNAs and Cancer: Mechanisms to Medicines ; 2015 Dec 4-7; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2016;76(6 Suppl):Abstract nr B38.</jats:p

Abstract B42: A two-lncRNA signature in serous exosomes serves as a new biomarker for colorectal cancer diagnosis

Abstract Background: Many studies have shown that long non-coding RNAs (lncRNAs) participate in the initiation and development of colorectal cancer (CRC). We previously reported that lncRNAs can be detected and are stable in serum. Exosomes (EXOs) are small (30 to 100 nm) membrane-bound particles that are released from normal, diseased, and neoplastic cells and are present in blood and other bodily fluids. EXOs contain a variety of molecules including signal peptides, mRNA, microRNA, and lipids. However, it is unclear if lncRNAs in serous EXOs represent a novel marker to detect CRC. Methods: Total RNA and serous EXO RNA were isolated from serum of 60 CRC patients, 60 age- and sex-matched healthy subjects. We measured 39 candidate cancer-associated lncRNAs by reverse transcription and quantitative polymerase chain reaction (RT-qPCR) in EXO RNA samples. The putative lncRNA markers identified in the training set (30 CRC patients and 30 healthy subjects) were verified in the test set (another 30 CRC patients and 30 healthy subjects). We further analyzed the putative lncRNA markers in serum total RNA samples. Results: In the training set, 16 lncRNAs were detectable in serous EXOs, and the expression levels of 7 lncRNAs were significantly different between healthy samples and CRC samples (p&amp;lt;0.05). Stepwise regression analysis showed that the combination of prostate cancer associated 3 (PCA3) and breast cancer anti-estrogen resistance 4 (BCAR4) provided the greatest predictive ability, with an AUC of 0.896 (95% CI: 0.794-0.921, p=0.029). Using the same serum samples, we compared the AUC values of these two lncRNAs in EXO with those in total RNA samples. The AUC values of our serous EXO two-lncRNA signature were markedly higher than those of serum signature (AUC=0.759, 95% CI: 0.696-0.853) for discriminating CRC patients from controls. The predictor was remarkably stable when applied to the test set comprising 30 CRC patients and 30 matched controls, with an AUC of 0.875. The AUC values of the serous EXO two-lncRNA signature for the test set were also markedly higher than those of serum signature (AUC=0.742 95% CI: 0.653-0.801) for discriminating CRC patients from controls. The results indicated that the two-lncRNA signature in serous EXO is an accurate biomarker for CRC diagnosis Conclusions: The PCA3 and BCAR4 signature in serous EXOs may facilitate the detection of CRC and serve as the basis for further studies of the clinical value of circulating lncRNAs in maintaining surveillance and forecasting prognosis. Citation Format: Peng Qi, Lei Dong, Wanrun Lin, Xiaoyan Zhou, Xiang Du. A two-lncRNA signature in serous exosomes serves as a new biomarker for colorectal cancer diagnosis. [abstract]. In: Proceedings of the AACR Special Conference on Noncoding RNAs and Cancer: Mechanisms to Medicines ; 2015 Dec 4-7; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2016;76(6 Suppl):Abstract nr B42.</jats:p

Correlation of immediate prevalence of cervical precancers and cancers with HPV genotype and age in women with atypical glandular cells cytology: A retrospective analysis of 369 cases

Background: This study aims to assess the immediate risk of cervical precancers and cancers in women with atypical glandular cells (AGC) cytology, based on high-risk human papillomavirus (hrHPV) genotypes and age. Methods: A retrospective analysis was conducted on 369 cases of AGC with immediate follow-up biopsy results, including 299 AGC-not otherwise specified (NOS) and 70 AGC-favor neoplastic (FN). Results: Among the 369 AGC cases, 127 tested positive for hrHPV (34.4%). The predominant high-risk type was other 11 genotypes (44.1%), followed by 16+ (29.1%), 18/45+ (26.0%), and 16 and 18/45 double-positive (0.79%). Precancers and cancers were detected in 30.4% (112 of 369) and 9.8% (36 of 369) of cases, respectively. The HPV-18/45+ group had notably higher adenocarcinoma in situ and adenocarcinoma (AIS+) prevalence compared to other 11 genotype groups (p \u3c.0001 and p =.001, respectively). The HPV-16+ group showed significantly higher high-grade cervical squamous epithelial lesion and squamous cell carcinoma prevalence than other 11 genotype groups (p \u3c.0001 and p =.017, respectively). Using 40-year cutoff, older women had significantly higher prevalence of abnormal glandular lesion+ lesions (17.6% vs. 7.6%, p =.005) and adenocarcinoma (AC) (12.4% vs. 2.5%, p =.001). Using 50-year cutoff, older women had higher prevalence of squamous cell carcinoma (SCC) (3.3% vs. 0.4%, p =.042) and AC (15.2% vs. 5.8%, p =.005). Subgroup analysis revealed that AGC-FN women showed more severe cervical pathology than AGC-NOS women (p \u3c.001). Conclusions: AGC women have a significantly increased risk of cervical precancerous lesions and cancer. HPV genotyping and patient age factors need to be taken into consideration in the clinical management process of AGC patients

Loss of tubal ciliated cells as a risk for “ovarian” or pelvic serous carcinoma

Abstract Background Recent advances suggest the fallopian tube as the main anatomic site for high-grade ovarian or pelvic serous carcinoma (O/PSC). Human fallopian tube is mainly lined by two cell types, secretory and ciliated cells. Large number of studies on the biologic role of tubal secretory cells in O/PSC have been performed in the last decade. However, the role of tubal ciliated cells in relation to the development of O/PSC has rarely been explored. The purpose of this study was to determine if change of the tubal ciliated cells shows difference in age and location and to examine their association with serous neoplasia. Methods Three groups (low-risk or benign control, high-risk, and O/PSC) of patients were age matched. The age data was stratified by 10-year intervals ranging from age 20 to older than 80. Ciliated cells from both tubal fimbria and ampulla segments were counted by microscopy and by tubulin immunohistochemical staining. The data was analyzed by standard contingency table, Poisson distribution methods, nonparametric Mann–Whitney U-tests and Spearman correlation analysis after age justification. Results The study revealed that the absolute number of tubal ciliated cells decreased significantly with age within each group. A reduction in ciliated cells within the fallopian tube remained a significant risk factor for serous neoplasia after age adjustment. A dramatic decrease of tubal ciliated cells in both tubal segments was identified in patients with high-risk and with O/PSC compared to those in the low-risk (benign control) group (p &lt; 0.001). Further, within the fimbria segment, a reduced number of tubal ciliated cells was more prevalent in the high-risk group when compared to those in O/PSC group. Conclusion Our findings suggest that reduced number of ciliated cells within the fallopian tube represents a hallmark of early serous carcinogenesis. Findings also support a relationship between loss of tubal ciliated cells and aging, the presence of high-risk factors, and co-existing ovarian or pelvic high-grade serous cancers. This represents an early study identifying the role of tubal ciliated cells in the process of high-grade O/PSC development.</jats:p

RAP80 is an independent prognosis biomarker for the outcome of patients with esophageal squamous cell carcinoma

Abstract Esophageal squamous cell carcinoma (ESCC) is the most popular pathology of esophageal cancer (EC) in China, especially in Henan province, mid-east of China. Presently, targeting DNA damage repair (DDR) factors is a promising approach for cancer therapy. Our group has been focusing on exploring the DDR factors overexpressed in ESCC tissues to provide potential targets for therapies for many years. RAP80/UIMC1 (ubiquitin interaction motif containing 1), one of those DDR factors we tested, was highly overexpressed in ESCC tissues compared with adjacent normal tissues. Moreover, the RAP80 mRNA level was validated to be an independent prognosis biomarker for the overall survival time of ESCC patients. The following biological assays revealed that it promoted cell proliferation both in vitro and in vivo, inhibited cell apoptosis at both early and late stages, and participated in G2/M checkpoint regulation. Even though studies have reported that ATM phosphorylates RAP80 at different serine sites upon DNA damage, the reversal regulation of RAP80 on the activity of ATM has never been investigated. In the study, mechanism explorations revealed that RAP80 positively regulated the ATM activity via proteasome–ubiquitination pathway to promote the transition of G2/M phase in cell cycle. By examining a number of E3 ubiquitination ligases (Ub) and deubiquitination (DUb) enzymes, we found that RAP80 positively regulated the stability of USP13 to promote cell proliferation of EC cells. Moreover, inhibition of RAP80 greatly sensitized EC cells to ATM inhibitor KU-55933, triggering a potential combination of RAP80 inhibitors and ATM inhibitors to enhance the therapeutic efficiency of ESCC patients for the clinicians

Engineering a red emission of copper nanocluster self-assembly architectures by employing aromatic thiols as capping ligands

Aromatic thiols are employed as capping ligands to prepare self-assembly architectures of Cu nanoclusters, which generate tunable aggregation-induced emission in the red region.</p

Fallopian tubal histogenesis of ovarian endometriosis—A study of folate receptor-alpha expression

BackgroundOvary is a common organ site involved by endometriosis. We previously found that fallopian tube may contribute to the histogenesis of ovarian endometriosis. The finding was novel and requires further studies. We addressed this issue by examining a differentially expressed gene folate receptor alpha (FOLR1) and its protein (FRA) in this study.ResultsA total of 144 tissue samples were studied. These included 32-paired tubal-endometrial-ovarian endometriosis samples (n = 96), 18 samples of ovarian endometriosis without corresponding fallopian tube or endometrium, and 30 ovarian tissue samples with ovarian surface epithelia but without endometriosis. Multiple comparisons among groups of ovarian endometriosis, normal fallopian tube and benign endometrium were performed. FOLR1 was highly expressed in the epithelia of fallopian tube and ovarian endometriosis, with paired endometrial samples showing a significantly lower level of expression. Similar differential studies for FRA protein were performed through Western blot and immunohistochemistry (IHC). The expression of folate receptor alpha at both mRNA and protein levels in the tissues (fallopian tube or ovarian endometriosis vs. the endometrium) were significantly different (p &amp;lt; 0.001). All ovarian surface mesothelial epithelia showed negative expression of FRA by IHC.ConclusionThe results further support that the fallopian tube may contribute to the development of ovarian endometriosis. Understanding the tubal contribution to ovarian endometriosis should ultimately contribute to ongoing investigative efforts aimed at identifying alternative ways to prevent and treat endometriosis. High level of FRA expression in the fallopian tube and endometriosis might be considered as potential tissue sites for targeted therapy.</jats:sec