Intraepithelial Cancer Research Articles

Cytology-based Screening for Anal Intraepithelial Neoplasia in Immunocompetent Brazilian Women with a History of High-Grade Cervical Intraepithelial Neoplasia or Cancer.

To determine the prevalence and possible variables associated with anal intraepithelial neoplasia and anal cancer in immunocompetent women with high-grade cervical intraepithelial neoplasia. A cross-sectional study involving immunocompetent women with a histological diagnosis of high-grade cervical intraepithelial neoplasia and cervical cancer, conducted between January 2016 and September 2020. All women underwent anal cytology and answered a questionnaire on characterization and potential risk factors. Women with altered cytology were submitted to anoscopy and biopsy. A total of 69 women were included in the study. Of these, 7 (10.1%) had abnormal anal cytology results: (high-grade lesion, atypical squamous cells of undetermined significance, and atypical squamous cells, cannot exclude high-grade lesions: 28,5% each; low grade lesion: 14,3%). Of the anoscopies, 3 (42.8%) showed alterations. Of the 2 (28,5% of all abnormal cytology results) biopsies performed, only 1 showed low-grade anal intraepithelial neoplasia. The average number of pregnancies, vaginal deliveries, and abortions was associated with abnormal anal cytology. However, the highest mean regarding the cesarean sections was associated with normal cytology. The prevalence of anal intraepithelial neoplasia was compatible with data from recent studies, especially those conducted in Brazil. Opportunistic screening for anal intraepithelial neoplasia in this high-risk population should be considered. Anal cytology is suitable for this purpose, due to its low cost and feasibility in public health services.

Effectiveness of Transvaginal Sonoelastography in Differentiation between Normal Cervix, Cervical Intraepithelial Neoplasia and Cancer Cervix

Effectiveness of Transvaginal Sonoelastography in Differentiation between Normal Cervix, Cervical Intraepithelial Neoplasia and Cancer Cervix

Vaccination against human papillomavirus after adjuvant therapy of cervical intraepithelial neoplasia

Background. The relevance of the problem is supported by the lack of effective secondary prevention measures against human papillomavirus (HPV)-associated cervical intraepithelial neoplasia (CIN) and cervical cancer. The only currently verified treatment for lower genital neoplasia is destructive therapy, with the recurrence rate of viral infection remaining quite high even after such interventions. The preventive role of HPV vaccine at pre-existing cervical lesions is actively debated worldwide, but the evidence remains scattered and contradictory. The present study surveys the scientific publications available that evaluate the efficacy of human papillomavirus vaccination as a secondary prevention measure against related cervical pathology.Objectives. An evidence survey on the efficacy of adjuvant vaccination against human papillomavirus in patients undergoing destructive therapy for cervical intraepithelial neoplasia.Methods. We have conducted a 10 year-depth review of scientific literature on the adjuvant human papillomavirus (HPV) vaccination efficacy for reducing the recurrence risk of precancerous cervical neoplasia. The results are presented in form of the difference of mean or pooled 95% confidence-interval odds ratios (OR; 95% CI). Statistical analyses were conducted with Review Manager v. 5.4 (The Cochrane Collaboration, 2020).Results. Thirteen Russian- and English-language studies were selected for final analysis. A total of 21,702 cervical dysplasia patients were included: 5,941 (27.4%) received a perioperative HPV vaccine, while 15,761 (72.6%) only had surgical treatment. Recurrences of CIN I (mild cervical intraepithelial neoplasia; OR 0.45, 95% CI 0.27–0.73; p = 0.001) and CIN II (moderate cervical intraepithelial neoplasia; OR 0.33, 95% CI 0.20–0.52; p <0.0001) were lower in the vaccinated vs. unvaccinated cohort.Conclusion. Adjuvant HPV vaccination associates with a lower risk of recurrence in cervical intraepithelial neoplasia. Further research is necessary to shed more light on the role of human papillomavirus vaccination as a secondary prevention measure against its associated lesions.

Abstract 806: Interrogating chromosomal instability in the earliest steps of ovarian cancer development

Abstract Ovarian Cancer is the deadliest gynecologic malignancy. Despite recent advances in cancer diagnostics, it is a cancer that is often detected late. After treatment with debulking surgery and chemotherapy, 80% of patients have tumor relapse. Early detection efforts have been largely unsuccessful and the only effective prevention strategy in high risk patients is surgical removal of the fallopian tube and ovaries. Careful pathologic studies of prophylactic risk reducing bilateral salpingo-oophorectomy (RRSO) has demonstrated that the origins of HGSC is likely within secretory cells of the fimbriae in the fallopian tube. This is based on numerous lines of correlative evidence including the identification of abnormal regions in the fallopian tube called serous tubal intraepithelial cancer (STIC) in 10-15% of patients undergoing RRSO, and the presence of STIC in half of patients undergoing debulking surgeries for HGSC. Finally, p53 mutations have been found to be identical in STIC and in the concurrent invasive disease indicating that HGSC and the STIC likely arise from the same clone. TCGA analysis has demonstrated that HCSC is a disease characterized by genomic and chromosomal instability harboring copy number alteration, including amplifications, deletions and more complex chromosomal rearrangements which impact multiple genes and pathways. Aberrantly segregating chromosomes that lag behind the spindle apparatus fail to become part of the primary nucleus (PN) and are instead enclosed by a self-assembled nuclear membrane, creating a structure called the micronucleus (MN). Spontaneous breakdown of the MN causes cytoplasmic DNA sensing and activation of the cGAS-STING pathway an important innate cellular defense mechanism to foreign cytoplasmic DNA. To understand the early genomic and signaling events in the development of HGSC, we are interrogating FFPE tissue from our archives from MSKCC patients who have undergone RRSO, hormone ablation surgeries or surgical debulking and were found to have benign fallopian tube, STIC lesions or invasive disease with STIC. We have performed a pilot experiment of 20 patient samples of different genetic backgrounds. Intriguingly, cGAS signaling was observed only rarely in the STICs indicating low rates of CIN in STICs, however we note that even in early invasive disease, we observe cGAS positivity. Moreover, we observe robust CD8+ T cell localization to early invasive disease. Taken together, these data are consistent with a model where CIN and subsequent cGAS/STING pathway activation drives the transition from pre-invasive to invasive disease with early immune recognition that is lost in invasive disease. We propose a model of ovarian cancer development where fallopian tube secretory transition from a p53 WT SCOUT to p53 mutant state STIL acquire a proliferative capacity forming a STIC and then develop CIN transitioning to invasive disease. Citation Format: Duaa Hassan Al-Rawi, Herman Chui, Sorhab Shah, Samuel Bakhoum. Interrogating chromosomal instability in the earliest steps of ovarian cancer development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 806.

16P The role of IL-8 gene polymorphism (845 T/C) in the development of cervical intraepithelial neoplasia

IL-8 gene polymorphisms are involved in the carcinogenesis of many malignant neoplasms, but their role in the development of cervical cancer (CC) has not been fully elucidated. The aim of the study was to evaluate the relationship between the presence of IL-8 gene polymorphism (845 T/C) and the development of severe cervical intraepithelial neoplasia (CIN) and cervical cancer.

Safety of the cyclin dependent kinase 9 (CDK9) inhibitor FIT039 for cervical intraepithelial neoplasia (CIN) 1 or 2 in a phase I/II trial.

3022 Background: Human papillomaviruses (HPVs) infect uterine cervical epithelial cells, leading to cervical intraepithelial neoplasia (CIN) and cervical cancer. However, there is no treatment for HPV infection in the uterine cervix prior to vaccination. We recently reported that cyclin dependent kinase 9 (CDK9) plays a critical role in viral RNA transcription of DNA viruses such as HPVs in host cells, and that FIT-039, a specific inhibitor of CDK9, suppresses the proliferation of several DNA viruses. Here, we evaluated the safety and antiviral effect of a FIT039-releasing vaginal tablet (FIT039CT) for CIN1 or 2 (CIN1/2). Methods: A multi-institutional, single-blind, placebo-controlled randomized phase I/II clinical trial involving 2 cohorts was designed to evaluate the safety of transvaginal FIT039CT for CIN1/2 as follows:. In the first cohort, 8 healthy women were randomized into FIT039CT (50mg/day or 100mg/day) or control group. In the second cohort, 14 women with a primary diagnosis of CIN1/2 were randomized into either FIT039CT (100mg/day) or control group. The primary endpoints were adverse events and plasma concentrations of FIT039. Results: 22 patients (8 volunteers, 11 CIN1 and 3 CIN2) were enrolled. There were no serious adverse events. Adverse events considered related to treatment were mild (vaginal discharge Grade 1: FIT039CT 16/17 women [94%] vs placebo 2/6 [33%]) and self-limiting in both cohorts. No patient discontinued this study due to adverse events. Maximum concentration (C max) and terminal elimination half-life (t 1/2) of serum FIT039 concentrations after single transvaginal treatment of FIT039CT were similar between the two doses as follows; C max (mean ± standard deviation) was 4.5 ± 0.5 ng/mL (50mg/day) and 4.4 ±1.4 ng/mL (100mg/day) at 6-7 hours; mean t1/2 was 14.8 ± 2.1 hours (50mg/day) and 12.1 ± 2.6 hours (100mg/day) hours. Conclusions: This study demonstrated the safety and validity of transvaginal FIT039CT once a day and may contribute to the development of an antiviral agent that can cure CIN1/2, and supports the design of the ongoing phase 2 clinical study. Clinical trial information: jRCT2051180201.

Alteration of Gene and miRNA Expression in Cervical Intraepithelial Neoplasia and Cervical Cancer

Background: Cervical cancer is one of the most common malignancies in women in terms of prevalence and mortality. Cervical cancer has some particularities that distinguish it from any other oncologic pathology: first, it is completely preventable by prompt detection of its precursor, cervical intraepithelial neoplasia (CIN); second, the Human Papillomavirus (HPV) infection is a known etiological agent; third, the mean age at diagnosis is much lower than in other oncologic conditions, as a consequence of the sexually-transmitted HPV. Methods: We evaluated the expression level of several long noncoding RNAs and a microRNA in samples from 30 patients with CIN, 9 with cervical cancer and 38 normal samples using qRT-PCR technology. Results: We observed higher expression levels for MEG3, DAPK1, MLH1 and MALAT1 in CIN samples than in normal samples, whereas TIMP3 and SOX1 had lower expression levels. For cancer samples, DAPK1, MLH1 and MALAT1 had higher expression, and MEG3, TIMP3 and SOX1 had lower expression when compared to normal samples. In the case of CIN versus cancer samples, only MEG3 gene showed a statistically significant difference. The expression of miR-205-5p was lower in both CIN and cancer samples compared to normal samples. Conclusion: Decreased MEG3 expression could be considered an alarm signal in the transition from a premalignant cervical lesion to invasive cancer, while altered expression levels of TIMP3, SOX1, MLH1, MALAT1 and miR-205-5p could serve as early biomarkers in the diagnosis of premalignant cervical lesions. Future studies, including a larger number of patients with CIN, will be of particular importance in validating these observations.

Implications of Persistent HPV52 and HPV58 Positivity for the Management of Cervical Lesions.

ObjectiveThis study aimed to compare the variability of HPV16/18/52/58 subtype infections in patients with different cervical lesions, to explore the guiding significance of persistent positive HPV subtypes 52 and 58 in the stratified management of cervical lesions, and to determine the appropriate management model.MethodThis study was conducted through a retrospective analysis of 244,218 patients who underwent HPV testing at the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University from September 2014 to December 2020 to examine the distribution of different types of HPV infection. From March 2015 to September 2017, 3,014 patients with known HPV underwent colposcopy to analyze high-risk HPV infection for different cervical lesions. Meanwhile, from September 2014 to December 2020, 1,616 patients positive for HPV16/18/52/58 alone with normal TCT who underwent colposcopy in our hospital were retrospectively analyzed for the occurrence of cervical and vulvovaginal lesions, with colposcopic biopsy pathology results serving as the gold standard for statistical analysis.ResultAnalysis of 244,218 patients who had HPV tested revealed that the top 3 high-risk HPV types were HPV52, HPV58, and HPV16. Further analysis of 3,014 patients showed that 78.04% of patients referred for colposcopy had HPV16/18/52/58 alone. Among high-grade squamous intraepithelial lesions (HSIL) and cervical cancer, the most common is HPV16, followed by HPV58 and then HPV52 (p < 0.05). A total of 1,616 patients with normal TCT who were referred for colposcopy due to HPV16/18/52/58 infection were further analyzed. Based on pathological findings in lesions of HSIL and CC, HPV16 is the most common, followed by HPV58 and then HPV18 (p < 0.05). In the 1,616 patients analyzed, high-grade vulvovaginal lesions were detected, with HPV58 being the most common, followed by HPV16 and then HPV52 (p < 0.05).Conclusion1. In patients with positive HPV58 alone and normal TCT, the indications for colposcopy may be relaxed, with particular attention paid to the possibility of vulvar and vaginal lesions.2. Patients with a positive HPV type 52 alone and normal TCT may be considered for a follow-up review and, if necessary, a colposcopy.3. The development of a more suitable HPV vaccine for the Asian population, such as HPV16/18/52/58, may better protect women’s health.

Comparison of conventional and liquid-based Pap smear methods in the diagnosis of precancerous cervical lesions

Cervical cancer is one of the five most common cancers among women. The present study aimed to compare conventional and liquid-based Pap smear methods in 2018 in Kerman, Iran. This was a cross-sectional study. Pap smear samples (conventional Pap smear (CN) and liquid-based cytology (LBC)) were collected from five health centres in Kerman. Samples were classified into two groups of liquid and CNs, and each group was classified into normal, abnormal (including, atypical squamous cells of undetermined significance (ASCUS+), atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H+), low-grade squamous intraepithelial lesion (LSIL+), high-grade squamous intraepithelial lesion (HSIL+) and cancer+) and unsatisfactory. Descriptive data analysis and chi-square/Fisher's exact test were performed in SPSS 20 (SPSS Inc., Chicago, IL). Totally, 31,513 women were screened by two CN and LBC. The mean age of subjects was 39.73 ± 9.58 years. The unsatisfactory smears (1.4% versus 0.02%; p=.001) and ASCUS+ (45.1% versus 39.4%; p=.007) were more reported by conventional tests, while LSIL+ (33.1% versus 38.9%; p=.005) was reported by the LBC test. Among women who were younger than 40 years, the CIN2+ that was found by LBC was significantly more than the CN method (37.4% versus 31.7, p=.04) and among older women (older than 40 years) the CIN1+ was significantly more as well (18% versus 13.6%, p=.05). The present study showed that, although LBC had many advantages, CN is still useful. Further studies are to be suggested as a clinical trial in another population with a large number of participants to compare the diagnosing methods of cervical cancers. Impact Statement What is already known on this subject? The conventional Pap smear (CN) and liquid-based cytology (LBC) methods are the most common methods for screening cervical cancers. Previously, in some studies, LBC was reported as a better method and in some studies, traditional method was preferred. Conflicting results were found in previous articles. What do the results of this study add? LBC compared to CN could find truer abnormal cases. The superiority of the LBC method is seen in cases such as reducing unsatisfactory cases, etc. LBC compared to CN could find more CIN+. Also, LBC could find more CIN1+ among women older than 40 years and more CIN+ ≥2 among younger women. What are the implications of these findings for clinical practice and/or further research? Although LBC has many advantages and it is easier, CN is still useful and both methods are suitable for cervical abnormality and cancer detection. CN cannot be declared an outdated method.

Evaluation of a prototype artificial life model to simulate the interactions between the human immune system, human papillomavirus type 16 and therapeutic vaccines

ObjectiveTo evaluate the artificial life model that simulates behaviours based on interactions occurring between the human immune system, the life cycle of human papillomavirus type 16, and certain types of therapeutic vaccines through virtual experiments on the constructed prototype. Materials and methodsWe developed the HPV16-ALIFE prototype using artificial life techniques to be used as a virtual laboratory. To evaluate the model, we analysed the responses of the simulated immune system under two circumstances: when a persistent infection is present that originates from HPV16 and causes lesions, and when the nanodevice releases a dose as part of a therapeutic vaccination strategy. Performing virtual trials means we can produce results for subsequent analysis and comparison with real-world clinical data. Results and conclusionsThe simulations enabled us to observe the proliferation of different cell populations considered in the model, levels of viral protein expression and cytokine secretion, transition stages of the developing infectious process-, low- and high-grade intraepithelial lesions, pre-cancer, and cancer status. The virtual patients who did not receive vaccines developed cancer status with peak rates of 18.86%. The patients who received a vaccination strategy reported a significant decrease in their cancer status, with peak rates varying between 0.48% and 3.12%, according to the specifications associated with each vaccine evaluated. The simulated immune system proved responsive to the activity of the virus and the vaccine.

Serum expression of Vascular Endothelial-Cadherin, CD44, Human High mobility group B1, Kallikrein 6 proteins in different stages of laryngeal intraepithelial lesions and early glottis cancer.

BackgroundThe study was designed to evaluate the potential validity and utility of selected molecular markers in serum samples from patients with specific stages of laryngeal intraepithelial lesions that could serve as diagnostic tools in differentiation of benign and dysplastic lesions from invasive pathologies.MethodsProspective study included 80 consecutive patients with vocal fold lesions treated at the single otorhinolaryngology centre. All participants had surgical resection of the lesion. Blood samples were collected from each patient before the surgery. Final diagnosis was confirmed on histopathological examination and included 39 (48.75%) non-dysplastic lesions, eight (10%) low-grade dysplasia, six (7.5%) high-grade dysplasia and 27 (33.75%) invasive cancers. The ELISA procedures were performed according to the manufacturer’s instruction. Individual serum concentration of selected proteins was reported in ng/ml: Vascular Endothelial-Cadherin Complex (VE-cad), CD44, Human High mobility group protein B1(HMGB1), Kallikrein 6.ResultsThe highest mean levels of HMGB1, KLK6 and VE-cad were detected in sera of patients with low-grade dysplasia (81.14, 24.33, 14.17 respectively). Soluble CD44 was the most elevated in patients with non-dysplastic lesions (2.49). The HMGB1, KLK6 and VE-cad serum levels were increasing from non-dysplastic to low-grade dysplasia and followed by the decrease for high-grade dysplasia and invasive cancer, however the differences were not significant (p-values 0.897, 0.354, 0.1 respectively). Patients’ serum had the highest CD44 concentration in non-dysplastic and low-grade dysplasia with the following decrease through high-grade dysplasia and invasive cancer. GERD symptomatic patients had higher levels of KLK6 and CD44 than other patients (p-value 0.06 and 0.084 respectively). There were no significant differences of biomarkers levels related to patients’ gender (p-value from 0.243 to 1) or smoking status (p-value from 0.22 to 0.706).ConclusionsVE-cad, HMGB1, CD44 and KLK6 did not prove to be reliable biomarkers implicating malignant potential within vocal fold hypertrophic intraepithelial lesions.

Genetic Polymorphisms in microRNA Genes Targeting PI3K/Akt Signal Pathway Modulate Cervical Cancer Susceptibility in a Chinese Population.

Polymorphisms in microRNA (miRNA) genes could influence the expression of miRNAs that regulate the PI3K/Akt signalling pathway and play crucial roles in cancer susceptibility. To investigate the association of single nucleotide polymorphisms (SNPs) in miRNA genes of PI3K/Akt with cervical intraepithelial neoplasia (CIN) and cervical cancer (CC), nine SNPs located in miRNA genes were selected for genotyping, and the association of these SNPs with CIN and CC risk was evaluated. A total of 1,402 participants were enrolled in the current study, including 698 healthy individuals in the control group, 431 patients with CC, and 273 patients with CIN. Nine SNPs in miRNA genes (rs107822 in miR-219a, rs10877887 in let-7i, rs2292832 in miR-149, rs353293 in miR-143, rs3746444 in miR-499, rs3803808 in miR-132, rs4078756 in miR-10b, rs629367 in let-7a, and rs7372209 in miR-26a) were genotyped using MassArray, and the association of these SNPs with CIN and CC were analysed. The results showed that the frequencies of rs107822 in miR-219a and rs2292832 in miR-149 were significantly different between the control and CC groups (p < 0.005). The C allele of rs107822 in miR-219a was associated with an increased risk of CC (OR = 1.29, 95%CI:1.09–1.54) whereas the C allele of rs2292832 in miR-149 was associated with a decreased risk of CC (OR = 0.77, 95%CI:0.64–0.92). The results of inheritance model analysis showed that the best-fit inheritance models for rs107822 and rs2292832 were log-additive. The 2CC + CT genotype of rs107822 could be a risk factor for CC when compared with the TT genotype (OR = 1.28, 95%CI:1.08–1.51). The 2CC + CT genotype of rs2292832 could be a protective factor against CC when compared with the TT genotype (OR = 0.76, 95%CI:0.64–0.92). However, no association of these SNPs with CIN was found in the current study. Our results suggest that rs107822 in the promoter region of miR-219a and rs2292832 in pre-miR-149 region are associated with the risk of CC.

Expression of p63 in Cervical Intraepithelial Neoplasia (CIN) and Cervical Cancer

Background: Cervical cancer is a major public health problem worldwide. Persistent infection with High-Risk Human Papilloma Virus (HPV) has been the main cause of squamous intraepithelial neoplasia which in turn leads to invasive squamous cell carcinoma. p63 is necessary for the activation of HPV, epithelial proliferation and differentiation. It also regulates the expression of certain cell cycle regulators. It has been reported that, from CIN I to CIN III, p63 expression increases progressively from basal layer to surface. In squamous cell carcinoma, it is expressed throughout the entire thickness of the tumor. Thereby it plays a significant role in diagnosing cervical premalignant and malignant lesions.&#x0D; Objective: To evaluate the relationship of p63 expression with different grades of CIN &amp; invasive SCC.&#x0D; Method: Total 86 paraffin embedded tissue blocks of histopathologically diagnosed cases of CIN and cervical cancer were evaluated by immunohistochemical staining for p63 expression. The study was performed in Sir Salimullah Medical College, Dhaka (from March, 2018 to February, 2020). Statistical analyses were carried out by using SPSS version 22 for Windows. A descriptive analysis was performed for all data. Observations were indicated by frequencies and percentages. Statistical significance was set at “p” value &lt;0.05.&#x0D; Results: Present study showed progressive increase in p63 expression from CIN I to CIN III from basal layer to surface. In invasive squamous cell carcinoma, higher expression of p63 was noted throughout the entire thickness of the tumor. No expression was seen in cervical adenocarcinoma and small cell carcinoma. In adenosquamous carcinoma only the area showing squamous differentiation revealed positive p63 expression. Statistically significant association of p63 expression was found with parity of patients and among grades of CIN .&#x0D; Conclusion: The results of this current study revealed that, p63 has significant association among different grades of CIN. It is also a useful marker in confirming a poorly differentiated squamous cell carcinoma &amp; predicting the progression of a squamous neoplastic lesion from cervical intraepithelial neoplasia to invasive squamous cell carcinoma. Moreover, it is useful to differentiate invasive squamous cell carcinoma from cervical adenocarcinoma.&#x0D; Sir Salimullah Med Coll J 2021; 29(2): 141-146

Observation of the cervical microbiome in the progression of cervical intraepithelial neoplasia

ObjectiveCervical microbial community in the cervical intraepithelial neoplasia and cervical cancer patients was analysed to study its composition, diversity and signalling pathways by high-throughput 16S rDNA sequencing,and the candidate genes associated with occurrence and progression of cervical intraepithelial neoplasia were screened out and the model was established to predict the evolution of cervical intraepithelial neoplasia malignant transformation from the cervical microbial genes aspect.MethodsCervical tissues of normal, cervical intraepithelial neoplasia and cervical cancer patients without receiving any treatment were collected. The correlation between candidate genes and cervical intraepithelial neoplasia progression was initially determined by analyzing the microbial flora. Real-time fluorescence quantitative PCR was used to detect the expression of candidate genes in different cervical tissues, ROC curve and logistic regression was used to analyse and predict the risk factors related to the occurrence and progression of cervical intraepithelial neoplasia. Finally, the early warning model of cervical intraepithelial neoplasia occurrence and progression is established.ResultsCervical tissues from normal, cervical intraepithelial neoplasia and cervical cancer patients were collected for microbial community high-throughput 16S rDNA sequencing. The analysis revealed five different pathways related to cervical intraepithelial neoplasia. 10 candidate genes were selected by further bioinformatics analysis and preliminary screening. Real time PCR, ROC curve and Logistic regression analysis showed that human papillomavirus infection, TCT severity, ABCG2, TDG, PCNA were independent risk factors for cervical intraepithelial neoplasia. We used these indicators to establish a random forest model. Seven models were built through different combinations. The model 4 (ABCG2 + PCNA + TDG) was the best early warning model for the occurrence and progression of CIN.ConclusionsA total of 5 differential pathways and 10 candidate genes related to occurrence and progression of cervical intraepithelial neoplasia were found in cervical microbial community. This study firstly identified the genes from cervical microbial community that play an important role in the occurrence and progression of cervical intraepithelial neoplasia. At the same time, the early warning model including ABCG2 + PCNA+TDG genes provided a new idea and target for clinical prediction and blocking the evolution of cervical intraepithelial neoplasia malignant transformation from the aspect of cervical microbiological related genes.

HPV and DNA Methylation Testing in Urine for Cervical Intraepithelial Neoplasia and Cervical Cancer Detection.

Biomarker detection in urine offers a potential solution to increase effectiveness of cervical cancer screening programs by attracting nonresponders. In this prospective study, the presence of high-risk human papillomavirus (hrHPV) DNA and the performance of DNA methylation analysis was determined for the detection of cervical cancer and high-grade cervical intraepithelial neoplasia (CIN2/3) in urine, and compared with paired cervicovaginal self-samples and clinician-taken cervical scrapes. A total of 587 samples were included from 113 women with cervical cancer, 92 women with CIN2/3, and 64 controls. Samples were tested for hrHPV DNA and five methylation markers. Univariate and multivariate logistic regression and leave-one-out cross-validation were used to determine the methylation marker performance for CIN3 and cervical cancer (CIN3+) detection in urine. Agreement between samples was determined using Cohen kappa statistics and the Spearman correlation coefficients. HrHPV presence was high in all sample types, 79% to 92%. Methylation levels of all markers in urine significantly increased with increasing severity of disease. The optimal marker panel (ASCL1/LHX8) resulted in an AUC of 0.84 for CIN3+ detection in urine, corresponding to an 86% sensitivity at a 70% predefined specificity. At this threshold 96% (109/113) of cervical cancers, 68% (46/64) of CIN3, and 58% (14/24) of CIN2 were detected. Between paired samples, a strong agreement for HPV16/18 genotyping and a fair to strong correlation for methylation was found. HrHPV DNA and DNA methylation testing in urine offers a promising solution to detect cervical cancer and CIN2/3 lesions, especially for women currently unreached by conventional screening methods.

Fas and Fas ligand are highly expressed in lymphocytes from cervical intraepithelial neoplasia and cervical cancer patients: A possible role for immune escaping.

Objective(s):Infection with high-risk human papillomavirus is required to develop cervical cancer. Some viruses modulate the Fas/FasL signaling to evade the immune response; the role of these molecules in cervical cancer is not clear. In this study, we measured the expression levels of Fas and FasL mRNA, soluble proteins, and cell surface proteins in peripheral blood mononuclear cells from patients with low- and high-grade squamous intraepithelial lesions and cervical cancer in relation to healthy women, to gain new insights into the role of Fas/FasL in cervical cancer development. Materials and Methods:Fas/FasL mRNA expression was measured in cervical tissues and peripheral blood mononuclear cells from patients and healthy subjects; serum soluble proteins Fas/FasL were measured by ELISA, and cell-surface protein expression was detected by flow cytometry. Results:Varying expression levels were found for both molecules. Cervical Fas and FasL mRNA expression was decreased in low- and high-grade lesions, but it was increased in cervical cancer cases. While, systemic Fas mRNA expression increased as malignity progressed; systemic FasL mRNA expression was increased in low- and high-grade lesions, but it was decreased in cancer patients. Soluble FasL levels decreased as lesions progressed, while soluble Fas levels increased. Finally, overexpression of Fas/FasL on the surface of peripheral blood mononuclear cells was found in patients with low-grade lesion with respect to healthy donors. Conclusion:Fas and FasL act as negative modulators of the immune response, probably by removing specific cytotoxic T lymphocytes against papillomavirus -infected cells and tumor cells.

PD-L1 as a Potential Therapeutic Target to Immune Checkpoint Blockade in Cervical Cancer: A Review of the Literature

Cervical cancer (CC) is still one of the most common malignant tumors in women worldwide. Malignant tumors are known for evading immune surveillance. In cervical lesions, the mechanism has received less attention. Evasion of immune surveillance through a variety of mechanisms; one of which is the immune checkpoint pathway has attracted interest because it can be therapeutically targeted, the programmed cell death 1 (PD-1) pathway that tumor cells use to block antitumoral immune responses. Increasing the prevalence of PD-L1 expression in up to 80% of cervical malignancies, there is a significant indication using immunotherapy to maintain the immune response against malignancies. The purpose of this literature review is to examine the potential role of immune checkpoint inhibitors in the treatment of cervical neoplastic lesions, by reviewing articles on cervical intraepithelial neoplasia (CIN) and cervical cancer that carried out histopathological and immunohistochemical examination PD-L1 as well as immunotherapy treatment with anti-PD-L1. There were 10 articles were found to be associated with anti-PD-1/PD-L1 immunotherapy. In conclusion, PD-L1 is a potential therapeutic target in cervical cancer

Clinical Significance of Detection of Human Papilloma Virus DNA and E6/E7 mRNA for Cervical Cancer Patients.

This study aimed to explore the clinical significance of detection of human papillomavirus (HPV) DNA and E6/E7 mRNA for cervical cancer patients. For this purpose, a total of 300 patients with cervical lesions who performed the colposcopy examination for the cervix between January 2018 and January 2020 were divided into three groups according to the level of cervical intraepithelial neoplasia (CIN): Low-level group (Group A, n = 101), high-level group (Group B, n = 149), cervical cancer group (Group C, n = 50) and gynecological inflammation group (Group D, n = 60). Tissue samples were collected from subjects above to perform the immunohistochemistry for E6/E7 protein and detection of HPV DNA and HPV E6/E7 mRNA. The results showed that HPV DNA copies and the positive rates of protein, DNA and mRNA of HPV E6/E7 in Groups A, B and C were significantly higher than those in Group D (all P<0.05), while no significant difference was identified in the comparison of the positive rate and copies of DNA among Group A, B and C (P> 0.05); in the Group A, B and C, patients had a higher positive rate of E6/E7 mRNA than those in the Group D (P<0.05), while in the Group B and C, the positive rate and copies of mRNA of HPV E6/E7 and HPV E6/E7 proteins were all higher than those in the Group A (P<0.05). In addition, the specificity of HPV E6/E7 mRNA was inferior to HPV DNA in the detection of the low-level intraepithelial neoplasia, but it performed better in the detection of the high-level intraepithelial neoplasia and cervical cancer. In general, HPV E6/E7 mRNA shows promising value in the detection of the development and progression of cervical cancer.

A novel cervical cancer screen-triage-treat demonstration project with HPV self-testing and thermal ablation for women in Malawi: Protocol for a single-arm prospective trial

Cervical cancer is the leading cause of cancer mortality among Malawian women, despite being preventable through screening and preventive therapy. In 2004, Malawi implemented a national screening program, using visual inspection with acetic acid (VIA) and cryotherapy, but its success has been limited due to equipment and human resources challenges. Since the development of that program, new technologies for screening and treatment that are less resource-intensive and more scalable have become available. GeneXpert systems provide fast, accurate HPV results and are increasingly available in low-income countries. Self-collection for human papillomavirus (HPV) testing is a validated method for screening and improves uptake. Thermal ablation provides an alternative ablative treatment that is simpler to use than cryotherapy and can be performed with portable devices. Meanwhile, urine HPV testing methods provide promising options for primary screening. We designed a single-arm prospective study to investigate a novel HPV screen-triage-treat strategy among 1250 women in Lilongwe, Malawi. Our proposed strategy consists of (1) Xpert HPV testing of self-collected samples, (2) VIA and colposcopy for HPV-positive women, and (3) thermal ablation for HPV-positive/ablation-eligible women. We will collect cervical biopsies, Pap smears, and endocervical samples to validate the HPV results and VIA/colposcopy findings against endpoints of high-grade cervical intraepithelial neoplasia or cancer (CIN2+). We will evaluate same-day completion of our algorithm, its performance in triaging women for treatment, and 24-week treatment efficacy of thermal ablation. We will also explore the performance of HPV and methylation tests in urine samples, as compared to provider- and self-collected cervicovaginal samples.

Co-Delivery of p53 Restored and E7 Targeted Nucleic Acids by Poly (Beta-Amino Ester) Complex Nanoparticles for the Treatment of HPV Related Cervical Lesions.

The p53 gene has the highest mutation frequency in tumors, and its inactivation can lead to malignant transformation, such as cell cycle arrest and apoptotic inhibition. Persistent high-risk human papillomavirus (HR-HPV) infection is the leading cause of cervical cancer. P53 was inactivated by HPV oncoprotein E6, promoting abnormal cell proliferation and carcinogenesis. To study the treatment of cervical intraepithelial neoplasia (CIN) and cervical cancer by restoring p53 expression and inactivating HPV oncoprotein, and to verify the effectiveness of nano drugs based on nucleic acid delivery in cancer treatment, we developed poly (beta-amino ester)537, to form biocompatible and degradable nanoparticles with plasmids (expressing p53 and targeting E7). In vitro and in vivo experiments show that nanoparticles have low toxicity and high transfection efficiency. Nanoparticles inhibited the growth of xenograft tumors and successfully reversed HPV transgenic mice’s cervical intraepithelial neoplasia. Our work suggests that the restoration of p53 expression and the inactivation of HPV16 E7 are essential for blocking the development of cervical cancer. This study provides new insights into the precise treatment of HPV-related cervical lesions.