Matthys H. Botha

Prognostic Relevance of Molecular Classification in Endometrial Cancer: Insights From a South African Cohort

PURPOSE Geographical and racial diversity may influence endometrial cancer (EC) prognosis, yet its impact remains underexplored. In South Africa (SA), the rising incidence of EC underscores the need to investigate potential biologic differences. Molecular classification of EC offers valuable prognostic insights that could help address disparities and improve care. This study evaluated the prevalence and prognostic significance of molecular subtypes in a South African high-intermediate and high-risk EC cohort. MATERIALS AND METHODS We included 133 patients with high-intermediate and high-risk EC diagnosed in SA between January 2017 and December 2021. Clinical, demographic (including self-identified race), and follow-up data were collected. Central pathology review assessed histotype, grade, lymphovascular space invasion, and International Federation of Gynecology and Obstetrics 2009 stage. Molecular subtyping followed the WHO 2020 algorithm using targeted next-generation sequencing and immunohistochemistry for p53, mismatch repair (MMR) proteins, and ER. Shallow whole-genome sequencing (sWGS) assessed genome-wide copy number alterations. RESULTS Among 131 patients with complete molecular classification, the most common subtype was p53-abnormal (p53abn, n = 71; 54.2%), followed by MMR-deficient (MMRd, n = 30; 22.9%), no specific molecular profile (NSMP, n = 21; 16.0%), and POLE -ultramutated ( POLE mut, n = 9; 6.9%). Nonendometrioid EC (NEEC) predominated (n = 82; 61.7%). High-grade endometrioid EC and NEEC were more frequent in non-White patients ( P = .030). Molecular subtypes were significantly associated with overall recurrence ( P = .029), with no recurrences in POLE mut ECs and the worst outcomes in p53abn ECs. sWGS revealed higher CN burdens in p53abn ECs, with recurrent focal alterations involving CCNE1 amplification and RB1 loss. CONCLUSION To our knowledge, this study is the first to demonstrate the prognostic value of EC molecular classification in a South African cohort. These findings support the global relevance of molecular EC subtyping. The urgent need for access to molecular diagnostics or cost-effective alternatives in resource-limited settings is highlighted.

Targeting treatment resistance in cervical cancer: A new avenue for senolytic therapies

Cervical cancer poses a significant global health challenge, particularly impacting women in economically developing nations. This disparity stems from a combination of factors, including inadequate screening infrastructure and resource limitations. However, the foremost contributor is the widespread lack of awareness and limited accessibility to Human Papillomavirus (HPV) vaccination, which is a key preventative measure against cervical cancer development. Despite advancements in cervical cancer prevention, treatment resistance remains a major hurdle in achieving improved patient outcomes. Cellular senescence, specifically the senescence-associated secretory phenotype (SASP) and its bidirectional relationship with the immune system, has been implicated in resistance to conventional cervical cancer chemotherapy treatments. The exact mechanisms by which this state of growth arrest and the associated changes in immune regulation contribute to cervical cancer progression and the associated drug resistance are not entirely understood. This underscores the necessity for innovative strategies to address the prevalence of treatment-resistant cervical cancer, with one promising avenue being the utilisation of senolytics. Senolytics are agents that have promising efficacy in clearing senescent cells from tumour tissues, however neither the utilisation of senolytics for addressing senescence-induced treatment resistance nor the potential integration of immunotherapy as senolytic agents in cervical cancer treatment has been explored to date. This review provides a concise overview of the mechanisms underlying senescence induction and the pivotal role of the immune system in this process. Additionally, it explores various senolytic approaches that hold significant potential for advancing cervical cancer research.

Cervical precancer thermal ablation versus LLETZ excision comparative efficacy study in WLWH (TALL Study): protocol for a randomised clinical trial in South Africa

Background Cervical cancer remains a significant global health concern and is the fourth most prevalent cancer among women. In South Africa, it is the leading cause of cancer-related deaths in women aged 15–44 years. The disease is typically preceded by persistent high-risk HPV infection, leading to cervical intraepithelial neoplasia and eventually cancer. Currently, in South Africa, management primarily involves excision, particularly through large loop excision of the transformation zone, which has associated risks and limitations. Thermal ablation is an alternative cost-effective treatment method, providing a straightforward approach to treatment, particularly advantageous in environments characterised by limited resources. The study aims to assess the efficacy, safety and patient experience of thermal ablation, providing valuable data for potential integration into South Africa’s cervical cancer prevention policies. Methods Randomised controlled trial in which 420 women living with HIV aged 30–60 years will be recruited from the Colposcopy Clinic at Tygerberg Hospital and will be followed up for a period of two years. The primary study endpoint is a test of cure that will be assessed by HPV genotyping, cervical cytology and histology at six month intervals. Other endpoints include the occurrence of adverse events. Ethics and dissemination The study protocol has been approved by the Health Research Ethics Committee of Stellenbosch University (Ethics Reference No: M20/11/035) and by the Western Cape Department of Health and Wellness via the National Health Research Database (WC_202109_016). All study procedures comply with the Declaration of Helsinki, South African Good Clinical Practice Guidelines and the Medical Research Council’s ethical guidelines. Trial results will be disseminated through peer-reviewed journals, national and international conference presentations and professional associations. A lay summary will be shared with the Community Advisory Board to guide community-level dissemination. Trial registration number Pan African Clinical Trial Registry: PACTR202504820339039.

High-grade cervical disease and cervical cancer in women aged 50 years and older compared with younger women: examining prevalence by HIV status in two large prospective cohorts in Botswana

Objectives International guidelines recommend cervical screening cessation at age 50 following two consecutive negative screens. However, many women aged 50 and older in low-income and middle-income countries (LMICs) have not had prior opportunity to screen. We examine the prevalence of cervical dysplasia and cervical cancer stage in Botswana women aged 50+ compared with 30–49, stratified by HIV status. Design Secondary analysis of data from two prospective cohort studies. Setting The screening cohort was recruited at health facilities in South East District. The cancer cohort was recruited from the primary public tertiary referral hospital and a private hospital in Gaborone, Botswana. Participants The screening cohort included 2570 women aged 30 and older recruited from February 2021 to August 2022. Screening eligibility included anyone with a cervix and without a prior history of cervical cancer. The cancer cohort included 1520 patients diagnosed with cervical cancer who sought care at the facilities where recruitment took place from January 2015 to December 2022. Primary and secondary outcome measures The prevalence of cervical intraepithelial neoplasia (CIN)2+ and cancer stage at diagnosis was compared across age groups, stratified by HIV status. Prevalence ratios were calculated for the association between age and CIN2+/CIN3+via log-binomial regression. Results The prevalence of CIN2+ was similar between 30–49 years old and 50+, both among women with HIV (WWH, 15.9% and 19.3%, respectively) and without HIV (13.3% and 10.4%, respectively). Similar findings were found when CIN3+ was used as the outcome. There were no statistically significant differences in prevalence ratios (PRs) across age groups for CIN2+ (adjusted PR (aPR) WWH 1.1 (95% CI 0.80 to 1.6); aPR HIV− 0.78 (95% CI 0.45 to 1.4) nor CIN3+ (aPR WWH 1.1 (95% CI 0.70 to 1.6); aPR HIV− 0.81 (95% CI 0.40 to 1.7)). Nearly half of cervical cancer diagnoses were made in women 50+; three-quarters of cases in women without HIV were diagnosed at 50+ years. Conclusions Our findings demonstrate the prevalence of high-grade cervical dysplasia and cervical cancer remains high beyond age 50 in both women with and without HIV in an LMIC context with high HIV prevalence. Screening women 50+ will allow treatment for cervical dysplasia and may provide early diagnosis of curable cervical cancer. These findings support the rapid introduction of high-performance cervical screening to increase access for women 50+. Trial registration number NCT04242823.

Inflammation and thrombotic risk in late-stage cervical cancer: An exploratory study of coagulation and cytokine profiles in a South African cohort

This exploratory study investigates the possible relationship between inflammation and thrombosis in cervical cancer patients in South Africa, highlighting the need for improved thrombotic risk profiling. Thromboelastography (TEG) was used to assess coagulation parameters in platelet-poor plasma (PPP) from a small cohort of late-stage (III and IV) cervical cancer patients (n = 19) and healthy controls (n = 15). Parameters assessed included clotting time, clot formation speed, and clot strength. A Luminex Multiplex assay was used to measure interferon-γ (IFN-γ), interleukin-1β (IL-1β), IL-6, vascular endothelial growth factor-A (VEGF-A), and tumour necrosis factor-α (TNF-α) in PPP. Haematological profiles were also evaluated. Cervical cancer patients displayed a significantly shortened clotting time (p = 0.0044) and increased clot strength (p = 0.0003), suggesting enhanced coagulation. IL-1β was notably elevated (p = 0.0200), consistent with an inflammatory environment. Higher lymphocyte, neutrophil, and platelet counts (p = 0.0162, 0.0420, and 0.0374, respectively) were observed, indicating a possible prothrombotic state. These findings suggest a potential link between inflammation and thrombosis in cervical cancer patients. However, due to this study's small sample size and exploratory nature, direct relationships between these factors have yet to be definitively established and remain speculative. Thrombotic risk profiling may still offer value in managing patients, but further investigation is required to confirm these preliminary observations.

Aptima HPV E6/E7 mRNA and cytology cross-sectional performance as primary screening tests for detection of high-grade cervical lesions in HIV positive and negative women in South Africa

Objective To assess the performance of APTIMA ® HPV E6/E7 mRNA assay (AHPV) with HPV 16 and 18/45 genotyping (AHPV-GT) and cytology in detecting cervical cancer and precancer in HIV positive and negative women in South Africa. Methods A multicentre cross-sectional study was performed in women aged 25–64 (n = 992) with cytology and AHPV with AHPV-GT reflex testing. All screen-positive and a random subset of screen-negative women were referred for colposcopy and biopsy. Results On cytology, low-grade squamous intraepithelial lesion (LSIL) or higher was found in 9.7% of HIV negative and 35.8% of HIV positive women. HPV mRNA positivity was 19.5% (4.4% HPV 16, 2.8% HPV 18/45, and 6.9% other high-risk HPV) in HIV negative women, compared to 45.8% (9.4% HPV 16, 9.7% HPV 18/45, and 27.6% other high-risk HPV) in HIV positive women . The prevalence of histological abnormalities in HIV negative vs HIV positive women was 24.3 vs 46.0% for cervical intraepithelial neoplasia (CIN) 2+, 10.2 vs 24.1% for CIN3+, and 1.4 vs 2.4% for invasive squamous cell carcinoma. AHPV sensitivity for detection of CIN3 + performed the best: 69.0% (95% confidence interval (CI) 56.8–81.1) in HIV negative vs 81.4% (95% CI 73.7–89.0) in HIV positive women, followed by ASCUS + (atypical squamous cells of undetermined significance) cytology: 58.6% (95% CI 45.7–71.6) vs 76.5% (95% CI 68.1–84.8). The best positive predictive value for CIN2 + was for AHPV-GT16, followed by AHPV-GT16,18/45 and cytology LSIL+: HIV-negative women 84.0% (95% CI 68.9–99.1); 76.9% (95% CI 63.3–90.6); 75.0% (95 CI% 61.2–88.9) and HIV-positive women 92.5% (95% CI 84.1–100); 86.8% (95% CI 79.1–94.6); 84.0% (95% CI 77.6–90.3). Conclusion Significantly more HPV infection and cytological/histological abnormalities and advanced disease were seen in HIV positive women. The lower than expected clinical sensitivities of all screening tests are comparable to HPV DNA sensitivities reported in similar populations. AHPV with AHPV-GT performed better than cytology as a screening and triage test.

Primary HPV-based Cervical Cancer Screening Algorithms in Botswana

Primary high-risk human papillomavirus (HPV) testing has become first line screening for cervical cancer in high-income countries. The feasibility of this approach in low- and middle-income countries (LMICs) is less clear, as is the role of HPV testing among women living with human immunodeficiency virus (HIV). The proposed study seeks to evaluate the accuracy of cervical cancer screening algorithms using primary HPV testing followed by various forms of visual evaluation, including visual inspection with acetic acid (VIA), colposcopy and automated visual evaluation (AVE) for the detection of high-grade cervical dysplasia, using histology as the gold standard. We will validate the AmpFire Assay for HPV self-sampling in our setting. We will determine safe screening intervals in women living with HIV (WLHIV) in an HPV-based cervical cancer screening program and compare triage strategies for positive HPV results at WHO recommended screening intervals for WLHIV. We also seek to understand in-depth the attitudes, acceptability and preferences regarding cervical cancer screening, HPV testing, and self-sampling, for women in Botswana through interviews of a sub-set of women recruited for the cervical cancer screening study. Finally, we will analyze the cost of two-stage cervical cancer screening algorithms using high-risk HPV testing in Botswana.