Within the United States, Hispanic women, especially those in Puerto Rico, face an increased risk of cervical cancer development. The objective of this study was to explore the cervical microbiota of Hispanic women at high risk of developing HPV-induced cervical dysplasia or with cervical dysplasia treated in Texas and Puerto Rico. Cervical swab samples were collected from 296 participants (N = 80 Texan Non-Hispanic White, N = 98 Texan Hispanic, and N = 118 Puerto Rican Hispanic) during each patient’s initial visit and subjected to 16S V4 rRNA gene sequencing for microbiome profiling. HPV types were grouped as HPV 16, other high-risk HPV types, and other using HPV genotyping. Among participants, 71% (N = 211) were classified as high-risk normal, and 29% (N = 85) had cervical dysplasia. HPV 16 was detected in 15% (N = 45), other high-risk HPV types in 33% (N = 98), while 52% (N = 152) of patients were classified as “other”. Comparative analysis of microbial community structures across locations revealed distinct compositions, with Texan Hispanic women showing higher alpha diversity for two alpha diversity metrics (Pielou evenness and Shannon Diversity Index). The prevalence of CSTs varied across locations and disease states, with CSTs III and IV-B being among the most common in the study cohorts. Overall, this descriptive study provides a better understanding of cervical microbiome in Hispanic women across multiple geographic locations, in order to guide future interventions.

Abstract Introduction: Oral cavity squamous cell carcinoma (OCSCC) and their precursor lesions, oral potentially malignant diseases (OPMD), arise within a complex mucosal microenvironment where the microbiome may contribute to carcinogenesis. In this cross-sectional observational study, we aimed to compare the bacterial communities of normal oral mucosa, OPMD, and OCSCC to determine whether specific compositional shifts or diversity patterns are associated with progressive disease. Methods: Eighty-one patients with OPMD (n=55) or OCSCC (n=26) undergoing clinical evaluation had site-matched swabs from lesions and contralateral clinically normal mucosa. Twelve patients without OPMD or any malignancy served as controls. Amplicon sequence variants (ASVs) generated from 16S rRNA gene amplicon sequencing data were taxonomically assigned to genus level. Alpha diversity (richness, Shannon, inverse Simpson) was compared within patients using paired Wilcoxon tests and across diagnostic groups using nonparametric tests. Beta diversity (Bray-Curtis) was assessed by principal coordinates analysis (PCoA) with PERMANOVA to estimate effect size (R²) and p values. Alluvial plots and Venn diagrams were generated to evaluate overlap between groups and inter and intra group diversity. Results: Cross-sectionally, alpha diversity did not differ significantly among controls, dysplasia lesions, and cancer lesions across all metrics. In contrast, PCoA of all groups showed modest but statistically significant compositional separation by disease status (PERMANOVA F=2.191, R²=0.052, p=0.001); within cancer patients, lesion versus contralateral normal sites showed a trend (F=1.159, R²=0.028, p=0.055), and within dysplasia patients, separation was minimal but nominally significant (F=0.647, R²=0.009, p=0.025). Overall, 49% of genera were shared among cancer lesions, dysplasia lesions, and controls, while 20% of genera were unique to dysplasia lesions and 10% were unique to cancer lesions. Lastly, both dysplasia and cancer lesions demonstrated higher intragroup Bay-Curtis distances compared with controls (FDR-adj. p<0.001), consistent with a progressive ecological diversification in malignant transformation. Conclusion: In this comparatively large, site-matched cross-sectional cohort spanning normal mucosa, OPMD, and OCSCC, oral carcinogenesis was characterized by shifts in beta diversity. Key findings include significantly increased evenness in dysplasia lesions relative to paired mucosa, small yet significant compositional differences by disease status (R² ≈ 5%), and an increase in unique ASVs within dyplasia and cancer lesions. These results support a model in which oral carcinogenesis is associated with subtle ecological reorganization rather than gross disruption of the microbiome and highlight candidate bacterial taxa for mechanistic and biomarker studies. Citation Format: Anastasios Maniakas, Zoey R. Neale, Jennifer Wargo, Jeffrey N. Myers, Nadim Ajami, Andrew G. Sikora, Lauren E. Colbert. Ecological shifts, not dysbiosis: Microbiome reorganization across oral precancer-cancer spectrum [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(8_Suppl):Abstract nr LB209.

OBJECTIVES:Bone metastases are common in advanced cancers. In patients with impending pathologic fractures, prophylactic fixation can improve quality of life. Postoperative radiotherapy (RT) is the standard of care for bone metastases; however, preoperative RT may be beneficial in some patients. We evaluated outcomes in patients treated with preoperative RT for bone metastases. METHODS:We performed a retrospective review of 10 patients with bone metastases treated with preoperative RT. Descriptive statistics were used to characterise the cohort, and the Kaplan-Meier method was used to estimate time to subsequent palliative RT treatment and overall survival. RESULTS:10 patients were included in the analysis. Preoperative RT was used for various reasons, including for continuation of systemic therapy (20%), to reduce the RT field (20%) and due to medical comorbidities delaying surgery (20%). The median time from completion of RT to surgery was 13 days (IQR 7-21). The majority of patients (90%) had no postoperative complications. No patients had radiographic evidence of local disease recurrence at a median of 13 months. CONCLUSIONS:Patients treated with preoperative RT do well with minimal operative complications and improvement in reported pain. A randomised clinical trial is warranted to compare outcomes for preoperative and postoperative RT for palliation of bone metastasis requiring orthopaedic intervention.

HPV-related cervical cancers pose challenges in murine models due to human-specific viral carcinogenesis. Patient-derived organoids (PDOs) enable to incorporate extracellular matrix and tumor microenvironment, and mirror human tumor features. Tumor and adjacent tissues were sampled via cytobrushes and resections, and then dissociated, and purified using a Lymphoprep gradient. One million cells were used for scRNA sequencing, while the rest formed PDOs. Baseline tumor and buccal swabs served as genomic controls. PDO identity was validated through immunofluorescent staining, bulk RNA sequencing, and whole exome sequencing (WES). Cell viability assays assessed PDO responses to bacterial metabolites, radiation, and chemotherapy. We established five PDO lines from cervical squamous cell carcinoma (CSCC) and cervical adenocarcinoma (CAC), including one normal PDO from CAC-adjacent tissue. Two primary cell lines were also developed: one from CAC and one from CSCC. Early cultures showed differential KI67 staining: CAC1101, CAC1446, and normal PDOs were positive, while CAC1237 and CSCC1188 were negative. Later cultures showed KI67 positivity across all lines. P63 staining was positive in early CAC1101, CAC1446-adjacent normal, and CSCC1188 PDOs but became negative in late CSCC1188. Only CAC1101 PDOs were PAX8-positive, and PAS staining was observed in all CAC-derived PDOs, including CAC1446-adjacent normal. Somatic mutation analysis revealed 22-46 mutations across most samples, except for Day-14 CAC1101 PDOs, which had 536 mutations. Day-21 CAC1101 PDOs showed 32.6%-54.5% similarity to their tumor swab. CSCC1188 samples exhibited consistent similarity with their tumor swabs (25.9%-40.0%) and within PDOs (37.0%-64.0%). Germline mutation analysis showed 1, 045-1, 918 mutations in most samples, except for Day-14 CAC1101 PDOs, which had 38 mutations. CSCC1188 samples showed higher similarity with their buccal swabs (71.3%-74.0%) and within PDOs (70.4%-78.4%) compared to CAC1101. Comparison of RNA sequencing data of CAC1101 and CSCC1188 showed samples from same patient sharing similar profiles. Cell viability assays indicated that bacterial metabolites increased resistance to radiation and cisplatin in CSCC1188-derived primary cells but had no effect on CAC1237-derived cells. We developed a workflow for generating and expanding PDOs from surgical and non-invasive cytobrush samples while preserving tumor characteristics. PDOs recapitulated the features of their original tumors, offering insights into tumor-microenvironment interactions and variable responses to chemoradiation therapy. Rui Wang, Dalissa Negrón-Figueroa, Xiaogang Wu, Bo Jiang, Barrett C. Lawson, Timothy A. Harris, Maura Gillison, Lauren Colbert. Characterization of cervical cancer patient derived organoid [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 37.

e17632 Background: Patients with advanced or recurrent endometrial cancer have a median progression-free survival (PFS) of 3-6 months and a 5-year survival rate of 17%, with limited therapeutic options. Recent studies indicate combination of chemotherapy and immunotherapy significantly increased PFS among patients with primary advanced or recurrent endometrial cancer, with a substantial benefit in mismatch repair-deficient (dMMR), microsatellite instability-high (MSI-H) population. Prior studies have shown reduced response to immunotherapy across different metastatic sites in various solid tumors, such as melanoma, colorectal and non-small cell lung cancer. This study explores the impact of metastatic sites on treatment outcomes in patients with advanced or recurrent endometrial cancer. Methods: In this single-center retrospective study of patients diagnosed with metastatic or recurrent endometrial cancer who received IO between 10/2019 and 12/2024, demographic characteristics, therapy received, site of metastatic disease (lung, liver, lymph node, brain), response to treatment were obtained from the medical records. Response was defined as partial or complete response and no response was defined as progression or stable disease. The association between site of metastatic disease and response was assessed using Fisher’s Exact Test. Results: We identified 21 patients with metastatic or recurrent endometrial cancer who received IO at MD Anderson Cancer Center. The majority identified as White (67%, n = 14) and Not Hispanic (60%, n = 12) with a median age of 68 (IQR: 61-71). Histologies include endometrioid (62%, n = 13), serous (33%, n = 7), clear cell (19%, n = 4) with 3 patients demonstrating multiple histologies. Patients received pembrolizumab and lenvatinib (67%, n = 14, median number of cycles 5.5) and pembrolizumab monotherapy (33%, n = 7, median number of cycles 9). The overall response to IO was stable disease (5%, n = 1), progression (38%, n = 8), partial response (38%, n = 8), and complete response (19%, n = 4). Patients received pembrolizumab and lenvatinib (67%, n = 14, median number of cycles 5.5) and pembrolizumab monotherapy (33%, n = 7, median number of cycles 9). The overall response to IO was stable disease (5%, n = 1), progression (38%, n = 8), partial response (38%, n = 8), and complete response (19%, n = 4). The odds ratios (OR) for overall response based on site of metastatic disease were liver (OR 1.77, p= 0.62), lung (OR 0.86, p= 1), lymph node (OR 1.04, p= 1), and bowel (OR not reported). Conclusions: This descriptive analysis identified patients with recurrent or advanced endometrial cancer with liver, lung, lymph node, and bowel metastases who received IO. Further retrospective data collection is underway to identify a possible role for site of disease, molecular signature, and response to IO.

PURPOSEThis study investigates the interplay between T-cell receptor (TCR) immune characteristics and microbiome profiles to explore the relationship between immune diversity and microbial composition in cervical samples from Ethiopia.METHODSCervical specimens were collected from patients at Tikur Anbessa Specialized Hospital in Addis Ababa, and rural Butajira, south-central Ethiopia. Patient data, including age, human papillomavirus status, pathology, and TCR immune characteristics, were analyzed with a focus on the interactions between TCR profiles and microbiome compositions in malignant samples.RESULTSThree distinct TCR profiles were identified: Group 1 (TCR active) exhibited features of active immune engagement, including high diversity, clonal expansion, and repertoire richness. Group 2 (TCR restricted) showed reduced TCR diversity and expansion, suggesting a restricted repertoire. Group 3 (TCR balanced) had moderate diversity and clonal activity. TCR repertoire groups were linked with microbial diversity, with Group 1 (TCR active) showing the highest number of microbes (high operational taxonomic units and microbial diversity). Maximum TCR clonal expansion positivity associated with microbial richness, while Group 3 (TCR balanced) was linked to reduced microbial alpha diversity. Taxonomic analysis revealed specific organisms enriched in TCR repertoire group.CONCLUSIONVariations in TCR profiles are linked to distinct microbial environments in cervical cancer with greater microbial richness in patients with greater maximum productive frequency. These findings underscore the interplay between TCR diversity, microbiome composition, and malignancy, offering insights into the potential implications for microbiome-targeted therapies and prognostic biomarkers in cervical cancer.

PURPOSECervical cancer remains a significant public health concern globally and particularly in sub-Saharan Africa, where high rates of HIV infection exacerbate cervical cancer incidence. Understanding the cervical microbiome and its role in cancer progression is essential, especially in regions where both cervical cancer incidence and HIV prevalence are high. This study aimed to characterize the cervical microbiome in women living with HIV (WLWH) and HIV-negative women with squamous cell carcinoma of the cervix in Botswana, compare the microbiome between before and after chemoradiation therapy (CRT) in WLWH, and assess the prognostic value of specific microbial taxa for overall survival (OS) in WLWH.PATIENTS AND METHODSCervical samples were collected from women with cervical cancer presenting to one hospital in 2018-2019. Patients' clinical data, including HIV status, were recorded. Microbial composition was analyzed using 16S rRNA gene sequencing. Microbiome diversity and composition were evaluated using alpha and beta diversity metrics. Differential microbial abundance was analyzed using linear discriminant analysis effect size. The association between microbial taxa and OS was explored using Cox proportional hazards regression.RESULTSWLWH (n = 42) had a significantly lower Pielou evenness index than HIV-negative women (n = 11; 0.6 v 0.7, P = .02), suggesting a more imbalanced microbiome in WLWH. WLWH had higher levels of Parvimonas and members of the Corynebacteriaceae and Micrococcaceae families, suggesting a shift toward a more pathogenic microbiome. In WLWH, CRT did not significantly alter overall microbial diversity. However, Lactobacillus and Sutterella were enriched before treatment, reflecting a less pathogenic microbiome, whereas Ruminococcus and Phascolarctobacterium and the families Caulobacterales and Flavobacteriia were enriched after treatment, reflecting microbial adaptations to the altered immune and treatment environment. Notably, higher levels of Flavobacteriia after CRT were independently associated with worse OS in WLWH.CONCLUSIONMicrobiome profiles differ between WLWH and HIV-negative women with cervical cancer in Botswana. The microbiome might have prognostic significance. Future research is needed to better understand the significance of the microbiota in cervical cancer progression and treatment outcomes and the potential role of microbiome-targeted interventions.

Purpose MRI-only based treatment planning workflow for gynecological brachytherapy can eliminate post-operative CT, optimize efficiency, reduce costs, and improve patient outcomes. The current study focusses the utility of commercially available needle MRI line markers (Orion, C4 Imaging, Houston, Texas) for Proguide needle digitization in hybrid gynecological applicators and in-house CISS MRI protocol specifically developed to maximize MR-marker signals . The study examines the dosimetric implications of MRI-only hybrid applicator reconstruction in comparison to conventional CT based reconstruction. Materials and Methods Seven clinical HDR -PDR brachytherapy cases were analyzed for gynecologic malignancies, utilizing two Geneva and five Venezia hybrid applicators, with three to six needles per case. Post-implant imaging included CT (Portable Airo® TruCT, Siemens Somatom Edge) with x-ray catheters and CT markers placed in intracavitary channels and needles, followed by CISS/Fiesta-C and T2-weighted MR images (T2 MRI) with intracavitary MRI line markers (Elekta, Stockholm, Sweden)and needle MRI line markers (C4 Orion) placed in intracavitary channels and needles. MRI was performed on a 1.5T scanner (Siemens Magnetom Sola) for two Venezia cases and on a 3T scanner (Siemens Magnetom Vida) for three Venezia and two Geneva cases. CT-based treatment planning was performed on Oncentra TPS (Elekta), using 3D applicator models for intracavitary channels and manual digitization for needles. The institutional workflow includes CT-MRI registration, with HR-CTV delineation using T2 MRI and OAR delineation on CT, followed by CT-based treatment planning in the Oncentra. Retrospective MRI-only based hybrid applicator reconstruction utilized intracavitary (Elekta) and needle (C4 Orion) MRI line markers in CISS/Fiesta-C MRI. No other changes of contouring and planning were performed. Both MRI line markers presented hyper signal intensity in CISS MRI sequence for 3D Applicator model and manual needle digitization (The dosimetric impact of MRI-only based hybrid-applicator reconstruction were quantified. Dosimetric evaluation compared the relative mean differences in HR-CTV dose coverage metrics (D90 and D98) and D2cc dose constraints for OAR’s between MRI-only based and conventional CT-based reconstruction plans. Results The mean percentage dose difference for HR-CTV D90 and D98 of MRI guided relative to CT-based planning was 0.6% ± 1.6% and -2% ± 3.9%, respectively. The mean percentage difference in dose for D2cc to the bladder, bowel, rectum, and sigmoid was 2.1% ± 6.5%, 3.5% ± 4.7%, -1.9% ± 3.6%, and -1.6% ± 4%, respectively. The p-values of the statistical test for HR-CTV D90 and D98 were 0.33 and 0.15, while for D2cc to the bladder, bowel, rectum, and sigmoid, the p-values were 0.45, 0.16, 0.31, and 0.50, respectively. The dosimetric differences between MRI-only and CT-based planning did not demonstrate statistical significance (p>0.05). Conclusions The Elekta MRI line markers and Orion MRI positive contrast line markers generated considerably hyper signal intensity in CISS MRI image for catheter visualization and facilitated MRI-only hybrid applicator reconstruction. MRI-only based hybrid applicator reconstruction plan yielded statistically comparable plans having dosimetrically equivalent to conventional CT-based plans. The current preliminary data results are demanded to be validated through statistically meaningful large datasets.

Abstract Objective Cervical cancer is a leading cause of cancer-related deaths in women worldwide, leading to over 340,000 deaths in 2022. The majority of cervical cancers are caused by the persistent infection of high-risk human papillomavirus (HPV). In the United States, Hispanic women are at increased risk of cervical cancer development and are more likely to die of the disease in certain geographical locations. Furthermore, prior research implicated an association between the vaginal microbiota and the risk of HPV infection and cervical dysplasia. This descriptive study aims to explore the vaginal microbiome profiles of Hispanic or Latino women at risk for HPV-induced cervical dysplasia or with cervical dysplasia residing in Houston and in Puerto Rico. MethodsCervical swab samples were collected from Hispanic patients in Houston IRB 2019-1059 (N = 93) and Puerto Rico IRB 1050411 (N = 118) during routine clinical visits on a prospective collection protocol. Swabs were acquired at the patient’s initial visit and subjected to 16S V4 rRNA gene sequencing for microbiome profiling. Through HPV genotyping via PCR, patients were categorized into three groups: having HPV 16, other high-risk HPV types (18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68), and “other”, comprised of low-risk HPV types, HPV negative, or unknown HPV status. Alpha and beta diversity were assessed using QIIME2. Microbiome composition was evaluated across different locations, HPV status, and disease status, followed by exploring alpha diversity between locations and disease status. The taxa species classification was performed using a custom classifier trained via a cervicovaginal-specific database. Samples were assigned to vaginal community state types (CSTs) using VALENCIA. Results Within the study cohort, 72% (N = 152) were classified as high-risk normal, and 28% (N = 59) had cervical dysplasia. HPV 16 was detected in 12% (N = 26), other high-risk HPV types in 33% (N = 70), while 55% (N = 115) of patients were classified as “other”. At the species level, the vaginal microbiota is dominated by Lactobacillus crispatus, Lactobacillus iners, and Trichomonas vaginalis as the top three abundant, regardless of the patient’s location, HPV status, and disease status. Alpha diversity between locations was 2.2 (1.4) for Houston and 1.6 (1.3) for Puerto Rico, as demonstrated by the Shannon Diversity Index. Alpha diversity was also observed between disease status, with a mean Shannon Diversity Index of 1.9 (1.4) for high-risk normal and 1.9 (1.1) for cervical dysplasia. The top three vaginal community state types were IV-B (32%), I (27%), and III (23%) for Houston, while the top three for Puerto Rico were III (34%), IV-B (31%), and I (21%). Conclusion The study examines the vaginal microbiome profiles of Hispanic women in Houston and Puerto Rico who are at risk for HPV-induced cervical dysplasia or already have cervical dysplasia. These findings provide insights into the vaginal microbiome composition, which plays a crucial role in cervical cancer development. Citation Format: D'Shaunique Walters, Molly El Alam, Timothy Harris, Tatiana Cisneros, Nadim Ajami, Jagannadha Sastry, Josefina Romaguera, Filipa Godoy-Vitorino, Stephanie Dorta-Estremera, Ann Klopp, Lauren Colbert. Descriptive study of the vaginal microbiome in Hispanic populations in Houston and Puerto Rico with high-risk normal and cervical dysplasia [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C082.

PurposeCervical cancer patients undergoing chemoradiation have high symptom burden. We performed an analysis of prospectively collected patient-reported outcomes(PROs) to determine characteristics predictive of poor treatment experience.MethodsBetween 2021-2023, we prospectively collected PROs from cervical cancer patients undergoing definitive chemoradiation. EORTC-QLQ-C30 and EORTC-QLQ-CX24 were completed at baseline(BL) and at the end of treatment(EOT). Poor treatment experience was defined as EOT poor health-related quality of life (HRQOL), low physical function, or significant overall symptom burden. Predictive factors analyzed included demographic, clinical, disease-specific factors, and baseline financial toxicity, depression, social function, and emotional function. ROC analysis provided appropriate predictive cut-off values. Univariable(UVA) and multivariable(MVA) linear regression analyses were performed.ResultsFourty-nine patients completed BL and EOT questionnaires. Median age was 43 (range, 18-85). Most patients (59%) had stage III disease. Baseline financial toxicity ≥66.7, depression ≥66.7, social function ≤50 and emotional function ≤58 on the EORTC linear transformed scale of 0-100 were significant predictors for poor treatment experience (p≤0.04) based on ROC analysis. On MVA poor BL social function was associated with reduced EOT HRQOL (β-9.3,_95%CI_-16.1_to_-2.6,_p<0.008), decreased physical function (β-24.4,_95%CI_-36.3_to_-12.6,_p<0.001), and high symptom burden_(β26.9,_95%CI_17.5_to_36.3,_p<0.001). Earlier disease stage predicted for decreased symptom burden_(β-6.7,_95%CI_-13.1_to_-0.3,_p=0.039). BL financial toxicity was a significant predictor on UVA (p=0.001-0.044) and showed a significant interaction term on MVA (p=0.024-0.041) for all three domains of poor treatment experience. Demographic and treatment-related factors were not predictive.ConclusionCervical cancer patients with poor baseline social function or high financial toxicity were at-risk for increased symptom burden and poor HRQOL. Screening for these factors provides an opportunity for early intervention to improve treatment experience.

For decades, chemoradiosensitization with checkpoint kinase inhibitors has been proposed but largely unexplored. A recent study reports that the novel ataxia telangiectasia and Rad3-related kinase inhibitor RP-3500 synergizes with radiation to control Atm-/- tumors in vivo. RP-3500 did not radiosensitize wild-type or Brca-1-deficient tumors, highlighting the need for a genotype-tailored approach. See related article by Ng et al., p. 5643.

Abstract Background: Cervical cancer patients undergoing CRT face a 40% mortality rate, and the genetic underpinnings of radiation response variability remain under-explored. Our laboratory has developed a noninvasive cervical swab biopsy method, complemented by a custom computational pipeline that facilitates longitudinal whole-exome sequencing (WES) from samples with minimal tumor purity. This investigation used the pipeline for the identification of persistent or clonally expanded genes during CRT. The aim is to identify genes and pathways associated with radiation resistance. Methods: Tumor swabs from 30 cervical cancer patients were collected at baseline (week 1) and CRT completion (week 5) paired with corresponding buccal samples. We performed whole exome sequencing, adjusting for calculated tumor purity with strict mutation calling using several tools. Results: Our analysis revealed genes in known DNA damage and repair (DDR) pathways, in addition to unique genes not previously associated with DDR and radiation response, including BAGE3, CGREF1, XRCC5, ATM, LINP1, etc. Future work will include validation of these identified genes in vitro radiation sensitivity screening and validation in large-scale datasets, such as The Cancer Genome Atlas, and network and pathway analysis. Discussion: This exploratory analysis suggests that serial sequencing during chemoradiation to identify novel radiation sensitization targets is feasible and further study is needed. Their recurring involvement in DDR pathways underscores their pivotal role in CRT resistance mechanisms. Conclusions: Through longitudinal WES, we've deepened the understanding of CRT resistance and flagged potential targets for improved radiosensitization strategies in cervical cancer. Future Directions: We are moving towards experimental validation of these genes in patient-derived organoids using CRISPR/Cas9 and CyTOF. Additionally, we'll be screening FDA-approved drugs to pinpoint effective radiosensitizers. A major upcoming effort is the development of a CRT resistance map, enhanced by machine learning, promising a transformative impact on cervical cancer treatment. Citation Format: Shafqat F. Ehsan, Rui Wang, Xiaogang Wu, Tatiana V. Karpinets, Julianna K. Bronk, Chiraag Kapadia, Xingzhi Song, Andrew M. Futreal, Ann H. Klopp, Tim Harris, Jianhua Zhang, Lauren E. Colbert. DDR pathway genes in CRT resistance: Insights from longitudinal whole exome sequencing in cervical cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: DNA Damage Repair: From Basic Science to Future Clinical Application; 2024 Jan 9-11; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2024;84(1 Suppl):Abstract nr A029.

Purpose The utilization of artificial intelligence in analyzing patient discussions on online platforms can uncover valuable experiential data that are often overlooked in structured surveys. Sentiment analysis, a branch of natural language processing (NLP), interprets and classifies emotions within text, offering insights into patient sentiments as positive, negative, or neutral. This study aimed to apply AI techniques to analyze the sentiments of posts on a cervical cancer-related online forum, specifically focusing on discussions related to brachytherapy. Materials/Methods Utilizing a Reddit Application Programing Interface, we extracted posts and comments from the subreddit r/cervicalcancer, focusing on discussions about brachytherapy between November 2020 and January 2024. We then processed the data in multiple steps including cleaning, lowercasing, removing illegible text, and tokenization. We analyzed the entries using RoBERTa (Robustly Optimized Bidirectional Encoder Representations from Transformers Pretraining Approach), a sophisticated pre-trained deep learning model, to refine and categorize sentiments. The model assessed the probabilities of the posts being positive, negative, or neutral. We further evaluated and categorized posts using pre-defined keyword tagging to uncover dominant topics within the conversations. These topics were modeled based on recently published literature related to the experiences of patients undergoing cervical brachytherapy. Results The analysis encompassed 879 out of 1,073 unique textual entries. Of these, overall sentiments were categorized as 40.1% positive, 30.1% negative, and 29.8% neutral. A specific focus on 'Bowel Domain’ discussions revealed a predominance of negative sentiments (51.2%)—the highest across all topics. Similarly, 'Urinary Domain' (46.8%), 'Fatigue' (42.4%), 'Anesthesia' (41.4%), and 'Pain' (43.4%) discussions largely reflected negative sentiments. In contrast, 'Physical Therapy' and 'Survivorship' discussions were predominantly positive, with 51.2% and 45.5% of posts, respectively. The sentiments on 'Sex' and 'Mental Health' related topics displayed a more balanced distribution between positive and negative perspectives. Conclusion This study demonstrates the value of using advanced AI models, such as sentiment analysis, to easily understand online patient discussions. These tools can bridge the gap between clinical insights and patient experiences, enhancing the feedback loop into clinical decisions, consent discussions, and patient education. Further research into the use of such models is necessary to fully leverage the insights they provide.

Herein, we present a protocol for culturing patient-derived organoids (PDOs) of cervical cancer that includes workflows for tumor biopsy/resection tissue and cytobrush-sampled cells. We describe steps for PDO culture initiation, including rinsing, gentle dissociation, Lymphoprep separation, and cell assessment, as well as seeding cells from surgical and cytobrush tissue digestion. We then provide guidance on PDO maintenance and passage and techniques for producing conditioned medium. Overall, this protocol serves as a valuable guide for establishing and maintaining cervical cancer PDOs.For complete details on the use and execution of this protocol, please refer to Colbert et al.1