High-risk Human Papillomavirus Types Research Articles

The combined finding of HPV 16, 18, or 45 and cytologic Atypical Glandular Cells (AGC) indicates a greatly elevated risk of in situ and invasive cervical adenocarcinoma

BackgroundCervical screening has not effectively controlled cervical adenocarcinoma (AC). Human papillomavirus (HPV) testing is recommended for cervical screening but the optimal management of HPV-positive individuals to prevent AC remains a question. Cytology and HPV typing are two triage options to predict the risk of AC. We combined two potential biomarkers (atypical glandular cell, AGC, cytology and HPV-types 16, 18, or 45) to assess their joint effect on detecting AC. MethodsKaiser Permanente Northern California (KPNC) used triennial co-testing with cytology and HPV testing (positive/negative) for routine cervical screening between 2003 and 2020. HPV typing of a sample of residual HPV test specimens was performed on a separate cohort selected from KPNC (Persistence and Progression, PaP, cohort). We compared risk of prevalent and incident histologic AC/AIS (adenocarcinoma in situ) associated with preceding combinations of cytologic results and HPV typing. Risk of squamous cell cancer (SCC)/cervical intraepithelial neoplasia grade 3 (CIN3) (SCC/CIN3) was also included for comparison. ResultsAmong HPV-positive individuals in PaP cohort, 99% of prevalent AC and 96% of AIS were linked to HPV-types 16, 18, or 45 (denoted HPV 16/18/45). Although rare (0.09% of screening population), the concurrent detection of HPV 16/18/45 with AGC cytology predicted a highly elevated relative risk of underlying histologic AC/AIS; the absolute risk of diagnosing AC/AIS was 12% and odds ratio (OR) was 1341 (95%CI:495–3630) compared to patients with other high-risk HPV types and normal cytology. Cumulatively (allowing non-concurrent results), approximately one-third of the AC/AIS cases ever had HPV 16/18/45 and AGC cytology (OR = 1785; 95%CI:872–3656). AGC was not as strongly associated with SCC/CIN3. ConclusionDetection of HPV 16/18/45 positivity elevates risk of adenocarcinoma, particularly if AGC cytology is also found.

Characterization of the oral microbiota among middle-aged men with and without human papillomavirus infection

ObjectivesTo characterize the oral microbiota among middle-aged men and identify differences between men with a prevalent oral high-risk (oncogenic) HPV infection and those without. Materials and MethodsThis was a case-control study nested within a prospective screening study for HPV-related cancers among middle-aged men. 16S rRNA sequencing was used to characterize the oral microbiota and the cobas HPV Test was used to detect presence of oral high-risk HPV types. We determined the overall composition of the oral microbiota and assessed differences in relative abundance of bacterial taxa as well as alpha and beta diversity among men with a prevalent oral high-risk HPV infection compared to men who were HPV-negative. ResultsAmong 13 high-risk HPV-positive and 30 HPV-negative men, we found significant differences in beta diversity but not alpha diversity. Fretibacterium, F0058, Kingella, Treponema, and Prevotella were more abundant among the high-risk HPV-positive men while Neisseria and Lactobacillus were more abundant among the HPV-negative men. ConclusionThis study adds to the evidence that the oral microbiota varies according to oral HPV infection status and may be associated with the natural history of oral HPV infection.

Detection of high-risk Human Papillomavirus in prostate cancer from a UK based population

Human papillomavirus (HPV) infection is one of the sexually transmitted diseases which have been implicated in the etiology of multiple cancers. To date, several studies have been conducted to evaluate the incidence of high-risk (HR) HPV in prostate cancer (PCa) which have generated widely conflicting data. Hence, this leaves a lack of awareness on the causal role of persistent HPV infection in the development of PCa. Although this has been investigated in a handful of countries, to the best of our knowledge, no prior studies have been conducted in the UK. In this study, polymerase chain reaction (PCR) and Sanger sequencing were implemented to analyze a total of 49 fresh prostate specimens (35 benign and 14 malignant specimens) for the presence of viral DNA of 12 HR-HPV types. Data obtained confirmed the presence of HR-HPV in 32.7% of analyzed benign and malignant prostate tissues with HPV 35 being identified as the most frequent type. Moreover, HR-HPV positivity rate was found to be higher in abnormal prostate tissues (adenocarcinoma and benign with prostatitis) compared those with normal prostate condition. Using immunohistochemistry, we have confirmed the expression of HPV E7 protein in prostate tissues positive for HPV DNA. This observation, the first reported from a UK population, suggests that the presence of HPV in prostate tissue is likely to be a related factor in the progression of certain cases of prostate cancer.

Protein-DNA Interactions Regulate Human Papillomavirus DNA Replication, Transcription, and Oncogenesis.

Human papillomavirus (HPV) is a group of alpha papillomaviruses that cause various illnesses, including cancer. There are more than 160 types of HPV, with many being "high-risk" types that have been clinically linked to cervical and other types of cancer. "Low-risk" types of HPV cause less severe conditions, such as genital warts. Over the past few decades, numerous studies have shed light on how HPV induces carcinogenesis. The HPV genome is a circular double-stranded DNA molecule that is approximately 8 kilobases in size. Replication of this genome is strictly regulated and requires two virus-encoded proteins, E1 and E2. E1 is a DNA helicase that is necessary for replisome assembly and replication of the HPV genome. On the other hand, E2 is responsible for initiating DNA replication and regulating the transcription of HPV-encoded genes, most importantly the E6 and E7 oncogenes. This article explores the genetic characteristics of high-risk HPV types, the roles of HPV-encoded proteins in HPV DNA replication, the regulation of transcription of E6 and E7 oncogenes, and the development of oncogenesis.

Splicing Characterization and Isoform Switch Events in Human Keratinocytes Carrying Oncogenes from High-Risk HPV-16 and Low-Risk HPV-84.

Infection of epithelial cells with high-risk HPV (HR-HPV) types, followed by expression of virus oncogenic proteins (E5, E6, and E7), leads to genomic imbalance, suppression of tumor inhibitors, and induction of oncogenes. Low-risk HPV (LR-HPV) may slow the rate at which cervical cancer spreads to an invasive stage since co-infection with LR-HPV is linked to a decreased risk of future invasive cancer than infection with HR-HPV alone. We then propose that cancer-progressing changes may be distinguished through identifying the functional differences between LR-HPV and HR-HPV. Lentiviral strategies were followed to establish HaCaT cells with constitutive expression of HPV oncogenes. RNAseq experiments were designed to analyze the transcriptome modulations caused by each of the E5, E6, and E7 oncogenes of HPV-16 and HPV-84 in HaCaT cells. We identified enhanced RNA degradation, spliceosome, and RNA polymerase pathways related to mRNA processing. ATTS (alternative transcription termination site) was discovered to be more prevalent in cells with HPV-16E5 than HPV-84E5. In HPV-16E6-infected cells, ATTS gain was significantly higher than ATTS loss. Cells with HPV-16E7 had more isoforms with intron retention (IR) than those with HPV-84E7. We identified switches in ADAM10, CLSPN, and RNPS1 that led to greater expression of the coding isoforms in HR-HPV. The results of this work highlight differences between LR-HPV and HR-HPV in mRNA processing. Moreover, crucial cervical cancer-related switch events were detected.

Interactions of the protein tyrosine phosphatase PTPN3 with viral and cellular partners through its PDZ domain: insights into structural determinants and phosphatase activity.

The human protein tyrosine phosphatase non-receptor type 3 (PTPN3) is a phosphatase containing a PDZ (PSD-95/Dlg/ZO-1) domain that has been found to play both tumor-suppressive and tumor-promoting roles in various cancers, despite limited knowledge of its cellular partners and signaling functions. Notably, the high-risk genital human papillomavirus (HPV) types 16 and 18 and the hepatitis B virus (HBV) target the PDZ domain of PTPN3 through PDZ-binding motifs (PBMs) in their E6 and HBc proteins respectively. This study focuses on the interactions between the PTPN3 PDZ domain (PTPN3-PDZ) and PBMs of viral and cellular protein partners. We solved the X-ray structures of complexes between PTPN3-PDZ and PBMs of E6 of HPV18 and the tumor necrosis factor-alpha converting enzyme (TACE). We provide new insights into key structural determinants of PBM recognition by PTPN3 by screening the selectivity of PTPN3-PDZ recognition of PBMs, and by comparing the PDZome binding profiles of PTPN3-recognized PBMs and the interactome of PTPN3-PDZ. The PDZ domain of PTPN3 was known to auto-inhibit the protein's phosphatase activity. We discovered that the linker connecting the PDZ and phosphatase domains is involved in this inhibition, and that the binding of PBMs does not impact this catalytic regulation. Overall, the study sheds light on the interactions and structural determinants of PTPN3 with its cellular and viral partners, as well as on the inhibitory role of its PDZ domain on its phosphatase activity.

A case of coexistent poorly differentiated adenosquamous carcinoma (glassy cell carcinoma), usual-type adenocarcinoma, and squamous cell carcinoma in situ of the cervix

Poorly differentiated adenosquamous carcinoma (glassy cell carcinoma) of the cervix is extremely rare, accounting for 1–2% of all cervical cancers. Herein, we report a case with coexistent poorly differentiated adenosquamous carcinoma (glassy cell carcinoma), “usual-type” adenocarcinoma, and squamous cell carcinoma in situ of the cervix. A female patient in her 60 s was referred to our hospital and diagnosed with poorly differentiated adenosquamous carcinoma based on cervical cytology and biopsy. The tumor was classified as clinical stage IB1 cervical cancer following magnetic resonance imaging; radical hysterectomy was performed. Histopathological examination revealed poorly differentiated adenosquamous carcinoma (glassy cell carcinoma), usual-type adenocarcinoma, and squamous cell carcinoma in situ, all coexisting. All carcinoma regions showed identical sizes to high-risk human papillomavirus (HPV) in fragment analysis. The patient is currently alive, without evidence of recurrence, 31 months post surgery. In this case, three different carcinomas coexisted. Fragment analysis of the patient’s HPV status suggested that all carcinomas were related to an infection with the same high-risk HPV type. To determine the precise mechanism of tumor development, i.e., whether the tumors were of the mixed or collision type, further studies are needed, including clonal analysis for the loss of heterozygosity pattern.

The prevention of cervical cancer

Cervical cancer screening concerns women between the ages of 25 and 65. It consists of the collection of cervical cells with a spatula by rubbing the cervix. The material was initially spread out and fixed on a glass slide. It was subsequently fixed in a liquid preservative with an automated spread on a thin-layer slide after centrifugation or filtration, a process called liquid cytology. Microscopic reading was facilitated by field selection using an automated pre-reading system. In July 2019, the French High Authority for Health (HAS) recommended to position DNA research of high-risk human papillomavirus types by PCR (HPV HR test) in first position after the age of 30. This approach is more sensitive than cytology in diagnosing a histological high-grade squamous intraepithelial lesion, and more effective in preventing invasive cancers. The HPV HR test, if positive, is followed by a cytological examination on the same sample to select patients requiring examination of the cervix by colposcopy. Vaccination against the nine most common types of HPV in girls and boys aged 11 to 14 years is the other part of the prevention of invasive cancer.

Prevalence of high-risk human papillomavirus infection and associated factors among women of reproductive age attending a rural teaching hospital in western Uganda

BackgroundHigh-risk HPV is considered a major risk factor for the development of cervical cancer, the most common malignancy among women in Uganda. However, there is a paucity of updated epidemiological data on the extent of the burden and factors associated with hr-HPV infection among women of reproductive age. The aim of this study was to determine the prevalence and genotype distribution of hr-HPV and associated factors among women of reproductive age attending a rural teaching hospital in western Uganda.MethodsWe conducted a cross-sectional study from April to June 2022. A total of 216 women of reproductive age attending the gynecological outpatient clinic were consecutively enrolled. Interviewer-administered questionnaires were used to collect participant characteristics, cervical specimens were collected by clinicians, and molecular HPV testing was performed using the Cepheid Xpert HPV DNA test. Descriptive statistics followed by binary logistic regression were conducted using SPSS version 22.ResultsThe prevalence of hr-HPV was 16.67%. Other hr-HPV types other than HPV 16 and 18 were predominant, with a prevalence of 10.6%; HPV 18/45 (2.31%), HPV 16 (0.46%), and 3.24% of the study participants had more than one hr-HPV genotype. On multivariate logistic regression, an HIV-positive status (aOR = 7.06, CI: 2.77–10.65, p = 0.007), having 3 or more sexual partners in life (aOR = 15.67, CI: 3.77–26.14, p = 0.008) and having an ongoing abnormal vaginal discharge (aOR = 5.37, CI: 2.51–11.49, p = 0.002) were found to be independently associated with hr-HPV infection.Conclusions and recommendationsThe magnitude of hr-HPV is still high compared to the global prevalence. HIV-positive women and those in multiple sexual relationships should be prioritized in cervical cancer screening programs. The presence of abnormal vaginal discharge in gynecology clinics should prompt HPV testing.

Modern approaches to cervical cancer screening (a review)

The aim – systematization of data from modern scientific sources and own knowledge and experience regarding the problems, advantages and prospects of the development of various methods of diagnosing pathological conditions of the cervix. The relevance of the problem of cervical cancer is beyond doubt, which is confirmed by the annual data of the official statistics and recommendations of WHO, European experts and relevant national recommendations. The analysis and generalization of information from the specialized literature regarding the assessment of problems and prospects for the development of leading methods of cervical screening in order to reduce the number of cervical cancer cases in the world is a priority task of the expert society. Currently, the most recommended approach to cervical screening is to combine HPV testing with cytology but separate testing for HPV is the most promising. Colposcopy remains the only confirmatory method of diagnosing cervical pathology, subject to mandatory cervical biopsy and histopathological examination of the biopsy. Standardization of classifications and terminology in the assessment of cytological, histopathological and colposcopic conclusions will allow to reach an understanding in the issues of optimal management tactics for patients with cervical pathology. Conclusions. Diagnosis of high-risk carcinogenic types of human papillomavirus is the most promising method of cervical screening in the near future. At the same time, at the moment, we consider it appropriate to use the cytology together with HPV testing, at least as a sorting method. Standardization of cytological/histopathological terminology should be done in accordance with the Bethesda 2014 system update.

Cervical human papillomavirus prevalence according to socioeconomic and demographic characteristics in a large Danish screening population.

The aim of this study was to investigate high-risk human papillomavirus (hrHPV) prevalence according to socioeconomic and demographic characteristics in a Danish screening population. We used data from HPV SCREEN DENMARK, which was an implementation study embedded into the routine cervical cancer screening programme. During 2017-2020, women aged 30-59 years screened in the Region of Southern Denmark were offered HPV testing or cytology. In the HPV group, liquid-based cytology samples were tested for 14 hrHPV types. We obtained registry information on socioeconomic and demographic characteristics and used log-binomial regression to estimate the prevalence ratio (PR) of hrHPV in three age groups (30-39, 40-49, 50-59 years), adjusting for age and marital status. We included 31,124 HPV unvaccinated women. In all age groups, the age-adjusted hrHPV prevalence was higher in women with basic versus higher education (e.g. age 30-39: 11.9% vs. 9.5%; PRage-adjusted=1.24; 95% confidence interval (CI): 1.02-1.50); women who were unemployed vs. employed (e.g. age 30-39: 11.6% vs. 10.4%; PRage-adjusted=1.11; 95% CI: 0.95-1.28); and in women with highest vs. lowest income (e.g. age 30-39: 11.6% vs. 9.5%, PRage-adjusted=1.18, 95% CI: 0.98-1.44). In models adjusted for marital status, these associations largely disappeared. We found slightly higher hrHPV prevalences in women with basic education, low income and unemployment. The differences largely disappeared when taking into account marital status as a potential proxy for sexual behaviour. Our findings support a need for targeted information on safe sexual practices and promoting socioeconomic equality in HPV vaccination and cervical cancer screening participation.

HPV Vaccination for Cervical Cancer Prevention in Switzerland

Human papillomavirus (HPV) is responsible for almost all cases of cervical cancer worldwide. It is also responsible for a variety of other cancers including penile; vaginal; vulvar; anal; and oropharyngeal cancers at the base of the tongue and tonsils. There are a very large number of these HPVs, which are classified into groups from high to low risk based on their oncogenic potential. Every year in Switzerland, over 260 women develop cervical cancer, and nearly 90 of them will die from the disease. Cervical cancer affects young women and is the fourth most common cancer in women between the ages of 20 and 49 years. Among the high-risk HPV types, HPV-16 and -18 are the most common and most carcinogenic ones. Together, these two HPV types are responsible for approximately 70% of cervical cancer cases in developed countries. HPV-6 and -11 are directly responsible for 90% of genital warts. There are two effective public health interventions to prevent this cancer: screening and vaccination. The present entry provides an overview of current literature in order to present these preventative approaches and consider their use within a Swiss context. It is hoped that, going forward, this will encourage the implementation and uptake of such interventions.

High prevalence and co-infection of high-risk Human Papillomavirus genotypes among unvaccinated young women from Paraguay.

Paraguay launched a human papillomavirus (HPV) vaccination program in 2013, so virological surveillance is important for measuring the impact of HPV vaccines. This study aimed to estimate the type-specific HPV frequency in unvaccinated sexually active women aged 18-25 years in the metropolitan area of Asuncion as a baseline for monitoring the HPV vaccination program. This study included 208 women, attending the Central Laboratory of Public Health between May 2020 and December 2021, were invited for testing through social networks and flyers at local health centers and higher education institutes. All participants who agreed to contribute to the study signed a free, prior, and informed consent form and answered a questionnaire that included basic demographic data and determining factors of HPV infection. Human papillomavirus detection and genotyping were conducted using the CLART HPV2 test (Genomica, Madrid, Spain) that allows the individual identification of 35 genotypes. 54.8% women were positive for any HPV type, with 42.3% positive for high-risk HPV (HR-HPV) types. Several factors were associated with HPV detection including the number of sexual partners, new sexual partners, non-use of condoms, and history of other sexual infections. Moreover, multiple infections were identified in 43.0% of the young women. We detected 29 different viral types present in both single and multiple infections. HPV-58 was the most commonly detected HPV type (14.9%), followed by HPV-16, HPV-51, and HPV-66 (12.3%). We estimated the prevalence of bivalent (16/18), quadrivalent (6/11/16/18), and nonavalent (6/11/16/18/31/33/45/52/58) vaccine types to be 8.2%, 13%, and 38%, respectively. These results reinforce the importance of surveillance studies and provide the first data regarding circulating HPV genotypes in the unvaccinated population in Paraguay, thus generating a baseline to compare future changes in the overall and type-specific HPV prevalence after HPV vaccination.

Associations of human papillomavirus genotypes and cervical vascular abnormality in a cohort of women underwent colposcopy, a retrospective study of 6716 patients.

Abnormal vessel patterns are specific signs in patients with early cervical abnormality and cervical cancer(CC) by colposcopy, but the impact of human papillomavirus (HPV) infections on abnormal vessel patterns remains unknown. A total of 6716 female patients with HPV infections or cytological abnormalities who underwent a colposcopy following abnormal CC screening results were included in the study. The final pathological diagnosis was confirmed to be the most severe pathological grade across cervical biopsy, endocervical canal curettage (ECC) and conization. Univariate and multivariate logistic regression analyses were used to investigate the association between HPV infections and abnormal vessel patterns, adjusting for age, gravidity and parity. There were 6124 normal vascular cases by colposcopy and 592 cases with cervical vascular abnormality. The prevalence of HPV infections was 4284 (70%) in normal patients, and the prevalence of HPV infections was 479 (80%) in cervical vascular abnormality patients. HPV high-risk type 16 infection alone increased the risk of cervical heteromorphic blood vessels (aOR=3.66, 95%CI: 2.54~5.27). HPV 16 and 33 alone (other than the commonly recognized subtype of 18) or coinfection of these two genotypes could increase the risk of cervical punctate vascular and cervical vascular mosaic features and abnormal cervical blood vessels. An increased risk of abnormal cervical lesions was observed for HPV 16 and 33 alone or combined in coinfection compared to the negative group. The risk of cervical vascular abnormality was increased 10-fold by coinfection with HPV 16 and 33 (aOR=10.67, 95% CI: 4.54~25.09, P<0.001). HPV 16, 33 alone or combined in coinfection were associated with an increased risk of lesions more advanced than high-grade squamous intraepithelial lesion (HSIL) when compared to the negative group. The risk of lesions more advanced than HSIL was up to 26-fold higher in the coinfection with HPV 16 and 33 group than in the negative group (aOR=26.23, 95%CI: 11.23~61.27, P<0.001). HPV16 and 33 are the most dangerous HPV genotypes correlated with abnormal vascular patterns. Combined HPV16 and HPV33 infection increases the risk of abnormal vascular patterns. Combined HPV16 and HPV33 infection increases the risk of developing HSIL+.

Abstract 6528: HPV status and epigenetic profiling of saliva from a cancer-free population

Abstract Identifying environmental factors and their relationship with epigenetic alterations is essential for understanding cancer, a leading cause of death. Currently, cancer screening and early detection development efforts are ongoing and epigenetics-based tests are one of the top choices as strategies. The understanding of how these alterations behave in cancer-free (CF) populations with well annotated data is essential. We randomly selected 601 saliva specimens from two Brazilian countrywide epidemiological studies POP-Brazil and SMESH. Extracted DNAs were sodium bisulfite converted and analyzed by Quantitative Methylation-Specific PCR (QMSP). Five genes previously reported as DNA methylation prone in cancer were selected (ITGA4, EDRNB, PAX5, p16, CCNA1). All participants were interviewed and provided sociodemographic and sexual behavior information as well as genital (gn) and oral biological samples. Variables were categorized and Fisher’s exact test was performed using SAS 9.4. Surprisingly, methylation in 3 cancer associated genes showed considerable frequencies in this CF group. DNA methylation was observed for EDNRB in 75/601 samples (12.5%), PAX5 in 55/601 (9.2%), and ITGA4 in 20/601 (3.3%). 12 samples displayed concomitant methylation on 2 of these 3 genes. p16 and CCNA1 methylation were only present in 1 sample each. Preliminary analyses suggest that specific genes methylation are observed with the presence of low (LR) and high-risk (HR) HPV types. EDNRB methylation groups with the presence of HR HPV type 16, 31, 39, 51, 58 and 59 from gn samples and LR types 6 and 42. ITGA4 methylation and HR HPV types 16 and 59 were detected in oral samples, and types 52, 56 and 68 in gn samples, as well as LR HPV types 40 and 42. In people with early onset of smoking, EDNRB and PAX5 seem to be more frequently methylated. The positivity of PAX5 was higher in participants who smoke, use alcohol and presented a positive diagnosis of syphilis and/or HIV. PAX5 methylation was more frequent in HR HPV 33, 58, 68 and 62 and LR type 6. Further analyses are ongoing. Our study shows methylation in cancer related genes in a CF population. The variables collected include some proven cancer risk factors and others still unknown, thus lead us to uncover directions to identify main cancer related players in a Brazilian population. The presence of cancer related alterations may indicate premalignant stages, exposure and lifestyle consequences, both at the same time or separately. In our pilot analysis, different types of HPV seem to correlate with epigenetic changes, this could bring attention to less studied HPV types and their role in carcinogenesis. Additional analyses of the current data and larger prospective, population-based studies, assessing the use of salivary DNA methylation in cancer-free and cancer groups of subjects are needed to explore the current findings and dissect the potential of saliva as a means of tracking cancer related changes. Citation Format: Ana Paula Muterle Varela, Dieila Giomo De Lima, Laura Palmieri, Juliana Comerlato, Fernando Hayashi Sant’Anna, Isabel C Bandeira, Marina Bessel, Pedro Isaacsson Velho, Eliana M. Wendland, Mariana Brait. HPV status and epigenetic profiling of saliva from a cancer-free population [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6528.

Abstract 3779: Evaluation of anti-cancer efficacy of lipid nanoparticles containing siRNA against HPV16 E6 and E7 combined with cisplatin in xenograft model of cervical cancer

Abstract Background: High-risk human papillomavirus (HPV) type 16 is the major etiological agent found in more than 60% of patients with cervical cancer and associated with cancer development and progression. Persistent expression HPV E6 and E7 oncoproteins are essential for the initiation and maintenance of cervical cancer. The therapeutic approaches targeting HPV E6 and E7 have been proved to be highly efficient in killing malignant cells. The suppression of HPV 16 E6/E7 expression by the short interfering RNA (siRNA) was more effective in combination therapy with cisplatin (CDDP) for cervical cancer. ENB101-LNP is an ionizable lipid nanoparticles (LNPs) encapsulating siRNA against E6/E7 of HPV 16 for delivery to the tumor cells. Methods: To investigate the anticancer efficacy and mechanism of ENB101-LNP combined with cisplatin in cervical cancer; the xenograft model was generated by the injection of CaSki cells subcutaneously into BALB/c nude mice. Mice were randomly divided into six groups: 1) blank control; 2) 1mg/kg ENB101-LNP; 3) 3mg/kg ENB101-LNP; 4) CDDP; 5) 1mg/kg ENB101-LNP-CDDP; 6) 3mg/kg ENB101-LNP-CDDP. ENB101-LNP was treated with intravenous injection three times weekly for 3 weeks and CDDP was treated with IP injection once a week for 3 weeks. Tumor growth curves were plotted to calculate the tumor inhibition rate. Mice were sacrificed one week after the last treatment and the tumor tissues were investigated using histopathology, immunohistochemical staining and western blotting. Results: Compared with the control and the other treatment groups, the tumor growth was significantly inhibited (P &amp;lt; 0.05). The tumor inhibition rate was 29.7% in 1mg/kg ENB101-LNP group, 29.6% in 3mg/kg ENB101-LNP group, 34.0% in CDDP group, 47.0% in 1mg/kg ENB101-LNP-CDDP group. At the dose of 3mg ENB101-LNP/kg combined with CDDP exhibited superior antitumor efficacy, the tumor inhibition rate was 68.8%. The successful ENB101-LNP mediated knockdown (with up to 80%) of HPV16 E6/E7 in tumors of both 1mg/kg and 3mg/kg groups was confirmed by RT-PCR. The ENB101-LNP increased p53 and p21 expression in tumors compared to control and CDDP alone groups and combination treatment with CDDP showed decreased expression of PD-L1 expression compared to CDDP alone group. The treatment of ENB101-LNP and combination with CDDP showed significant increase in apoptotic cells compared to control and single-agent groups respectively. Conclusions: The ENB101-LNP inhibiting E6 and E7 of HPV 16 in combination with CDDP showed promising anticancer activity in the cervical cancer cells xenograft mouse model. Citation Format: Sung Wan Kang, Shin-Wha Lee, Ji-young Lee, Ok Ju Kang, Hyejeong Kim, Yisak Kim, Yong-Man Kim. Evaluation of anti-cancer efficacy of lipid nanoparticles containing siRNA against HPV16 E6 and E7 combined with cisplatin in xenograft model of cervical cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3779.

Abstract 6062: Viral genome structure correlates with patterns of somatic alteration in human papillomavirus associated head and neck squamous cell carcinoma

Abstract Integration of viral DNA into the host genome has been implicated in human papillomavirus (HPV) associated tumorigenesis, and studies in HPV associated head and neck squamous cell carcinoma (HNSCC) attempting to classify viral integration status have reported associations among HPV status, viral load, and overall survival. However, few genomic studies have correlated HPV structure with commonly occurring somatic mutations in HNSCC. Here, we conducted hybrid-capture DNA sequencing using a panel of over 800 cancer related genomic targets, as well as probes specific for the full HPV16 and HPV18 genomes, in a novel cohort of 525 HNSCC cases. Paired reads were aligned to a human reference containing complete viral genomes, including all high-risk HPV types, which allowed for robust identification of HPV(+) tumors and specific HPV type. Variant calling and filtering, followed by pathway analysis of significantly mutated genes, defined the catalog of somatic driver alterations in HNSCC. Viral genome structure, integration status, and viral load were determined via initial copy number analysis of HPV-specific reads, followed by identification of viral/human breakpoints and indels with variant calling algorithms. Manual review was required in some cases due to variability in read coverage and alignment to HPV genome. We identified 252 HPV(+) tumors, the majority of which were HPV16 (n=228, 90%). Analysis of HPV16(+) tumors revealed five classes with variable viral loads: pure episome (EPI, n=54), episomal rearranged (EPI-R, n=51), episomal chimeric (EPI-C, n=22), integrated (INT, n=92), and integrated deleted (INT-A, n=12). Integrated tumors had at least one integration event (INT) and a few exhibited recurrent deletions in the viral genome (INT-A). Episomal samples had either complete HPV genomes (EPI), circular genomes with one or more indels (EPI-R), or circular genomes with HPV and human DNA (EPI-C). Interestingly, PIK3CA, the most frequently mutated gene in HPV(+) HNSCC, was mutated at a higher frequency in all integrated samples compared to all episomal types (44% vs 23%, p=0.005). We also noticed more deleterious mutations in innate immunity genes (HLA-A/B, B2M, TRAF3, BIRC3) in the episomal samples (16% vs 8%), which may allow cells with intact HPV genomes to evade anti-viral immune responses. Additionally, mutations in oxidative stress response genes that lead to the constitutive activation of NRF2 (NFE2L2, CUL3, KEAP1) were more frequent in integrated samples (11% vs 4%), which could confer radiotherapy resistance and a worse prognosis. Overall, using DNA sequencing and integrative genomic analysis in the largest cohort of HPV(+) HNSCC to date, we developed a novel classification method based on viral genome structure which associates with host somatic alterations that may contribute to variable outcomes in HNSCCs. Citation Format: Jeremiah Ray Holt, Xiaobei Zhao, Hyo Young Choi, Paul Little, Angela L. Mazul, Benjamin Wahle, Vonn Walter, Jose P. Zevallos, David Neil Hayes. Viral genome structure correlates with patterns of somatic alteration in human papillomavirus associated head and neck squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6062.

Infection with human cytomegalovirus, Epstein-Barr virus, and high-risk types 16 and 18 of human papillomavirus in EGFR-mutated lung adenocarcinoma

To assess the frequency of human cytomegalovirus (HCMV), Epstein-Barr virus (EBV), and high-risk types of human papillomavirus (HPV16 and HPV18) infections in lung adenocarcinoma samples.Lung adenocarcinoma cytological smears and their DNA isolates were obtained from patients hospitalized at the Department for Lung Diseases Jordanovac, Zagreb, in 2016 and 2017. Overall, 67 lung adenocarcinoma samples were examined: 34 with epidermal growth factor receptor gene (EGFR) mutations and 33 without EGFR mutations. The EGFR mutation status and virus presence were assessed with a polymerase chain reaction, and random samples were additionally tested for EBV with Sanger sequencing. HCMV, EBV, HPV16, and HPV18 infections were evaluated in relation to EGFR mutation, smoking status, and sex. A meta-analysis of available data about HPV infection in non-small cell lung cancer was performed.More frequent HCMV, EBV, HPV16, and HPV18 infections were observed in lung adenocarcinoma samples with EGFR mutations than in samples without these mutations. Coinfection of the investigated viruses was observed only in lung adenocarcinoma samples with mutated EGFR. In the group with EGFR mutations, smoking was significantly associated with HPV16 infection. The meta-analysis showed that non-small cell lung cancer patients with EGFR mutations had a higher odds of HPV infection.HCMV, EBV, and high-risk HPV infections are more frequent in EGFR-mutated lung adenocarcinomas, which indicates a possible viral impact on the etiology of this lung cancer subtype.

CLINICAL STUDY OF PREVALENCE OF HUMAN PAPILLOMA VIRUS (HPV) BY ELISA METHOD IN FEMALES ATTENDING GYNAECOLOGY OPD

Background: Women with normal cervical cytology, who are infected with high-risk HPV type, have an approximately 100-fold increased risk of developing cervical cancer compared with uninfected women. Present study was conducted to study seroprevalence of Human Papilloma Virus by ELISA method in female patients attending in Gynecology OPD of in a tertiary care hospital. Present study was cross- Material and Methods: sectional study, observational study, conducted female patients clinically suspected of having HPV infections like bleeding per vagina, irregular menstruation, pain in lower abdomen or post-coital bleeding or had any abnormal ndings on per vaginal examination. Among 160 Results: patients, majority of were from age group 31-40 years (39.38%), married (88.75 %), married at 18-20 years age (54.36 %), had parity of 2-4 (83.75%). All females had gynecological problems followed by women who were smoker ( 33.13 %), suffering from STD's (29.37 %), had alcohol drinking habit (25 %), had multiple sexual partner (5 %) and family history of cervical cancer (2.5 %). The patients who used homemade napkins had a highest seroprevalence of HPV 68.18% followed by patients who are suffering from STD's (63.83 %), had a family history of cervical cancer (50 %), women who used to smoke (37.7 %), drink (25 %), had multiple sex partners (25 %) and the least number seropositivity was among patient who had gynecological problems 23.12%. The sero-prevalence among patients for HPV was 23.12%. Conclusion: The prevalence increased with early marriages, high parity, and middle socio-economic status group. So, our study will guide for planning and implementation of governmental HPV infection control measures and the consequences of infection around the world. ELISA method is an important diagnostic method in future in HPV infection detection as it has several advantages over other existing diagnostic methods in respect of early detection, reliability, cost effectiveness and patients compliance as blood can be easily accessible.

HPV genotyping of cervical histologic specimens of 61, 422 patients from the largest women hospital in China.

We investigated HPV genotypes in a large cohort of patients with definitive cervical histologic diagnosis. HPV testing was performed by real-time PCR assay, including 18 high-risk HPV (hrHPV) and 3 low-risk HPV (lrHPV). Totally 61,422 patients with documented HPV genotyping results within 6 months before cervical histologic diagnoses were included. HrHPV positive rate was 55.1% among all tested cases with the highest in adenosquamous carcinoma (94.1%), followed by squamous cell carcinoma (SCC) (93.7%), cervical intraepithelial neoplasia 2/3 (CIN2/3) (92.8%). HrHPV positive rates were significantly higher in high-grade squamous lesions than in those in glandular lesions. HPV16 was the most common genotype followed by HPV52 and HPV58 in CIN2/3. The most frequent hrHPV genotype in adenocarcinoma in situ (AIS) was HPV18, followed by HPV16, HPV45 and HPV52. In SCC cases, HPV16 was the most common type followed by HPV58, HPV52, HPV18 and HPV33. However, HPV18 showed significantly higher prevalence in adenocarcinoma and adenosquamous carcinoma than in that in SCC. Theoretically, the protective rates of 2/4-valent and 9-valent vaccine were 69.1% and 85.8% for cervical cancers. The prevalence of HPV genotypes in Chinese population was different from that in Western population. Some hrHPV types were identified in cervical precancerous lesions and cancers, which are not included in current HPV vaccines. These data provide baseline knowledge for future HPV vaccine development.