Intraepithelial Neoplasia Lesions Research Articles

Cytokine CCL9 Mediates Oncogenic KRAS-Induced Pancreatic Acinar-to-Ductal Metaplasia by Promoting Reactive Oxygen Species and Metalloproteinases.

Pancreatic ductal adenocarcinoma (PDAC) can originate from acinar-to-ductal metaplasia (ADM). Pancreatic acini harboring oncogenic Kras mutations are transdifferentiated to a duct-like phenotype that further progresses to become pancreatic intraepithelial neoplasia (PanIN) lesions, giving rise to PDAC. Although ADM formation is frequently observed in KrasG12D transgenic mouse models of PDAC, the exact mechanisms of how oncogenic KrasG12D regulates this process remain an enigma. Herein, we revealed a new downstream target of oncogenic Kras, cytokine CCL9, during ADM formation. Higher levels of CCL9 and its receptors, CCR1 and CCR3, were detected in ADM regions of the pancreas in p48cre:KrasG12D mice and human PDAC patients. Knockdown of CCL9 in KrasG12D-expressed pancreatic acini reduced KrasG12D-induced ADM in a 3D organoid culture system. Moreover, exogenously added recombinant CCL9 and overexpression of CCL9 in primary pancreatic acini induced pancreatic ADM. We also showed that, functioning as a downstream target of KrasG12D, CCL9 promoted pancreatic ADM through upregulation of the intracellular levels of reactive oxygen species (ROS) and metalloproteinases (MMPs), including MMP14, MMP3 and MMP2. Blockade of MMPs via its generic inhibitor GM6001 or knockdown of specific MMP such as MMP14 and MMP3 decreased CCL9-induced pancreatic ADM. In p48cre:KrasG12D transgenic mice, blockade of CCL9 through its specific neutralizing antibody attenuated pancreatic ADM structures and PanIN lesion formation. Furthermore, it also diminished infiltrating macrophages and expression of MMP14, MMP3 and MMP2 in the ADM areas. Altogether, our results provide novel mechanistic insight into how oncogenic Kras enhances pancreatic ADM through its new downstream target molecule, CCL9, to initiate PDAC.

Abstract LB324: STING protein expression is increased in intermediate luminal cells in proliferative inflammatory atrophy of the prostate (PIA) and is absent in the majority of primary prostatic adenocarcinomas

Abstract Unresolved chronic inflammation has been postulated to drive prostate cancer development. We have proposed that proliferative inflammatory atrophy (PIA) can lead to the development of preneoplastic high grade prostatic intraepithelial neoplasia (PIN) lesions and/or directly to adenocarcinoma at times. Additionally, some PIA cells harbor molecular alterations seen in PIN and invasive carcinoma lesions, with a low frequency of PIA cells showing MYC over-expression, partial promoter hypermethylation and silencing of GSTP1, and formation of ETS gene fusions. PIA appears to result from inflammatory injury and regeneration in which proliferative and atrophic epithelial cells have an intermediate cell phenotype with both luminal and basal cell features, including basal cell markers, keratin 5 and BCL2, as well as varying levels of prostate luminal enriched genes such as androgen receptor, PSA and NKX3.1. Also, a number of gene products that are apparently stress induced, which are not expressed in normal prostate are induced in at least a subset of intermediate luminal cells in PIA, including GSTA1, PTGS2, LTF and PIGR (PMIDs:17384581, 37550827). Stimulator of interferon genes (STING), encoded by TMEM173, is a multipass transmembrane ER associated protein, and is a critical hub for innate responses directed against numerous bacterial, viral and parasitic pathogens, and is important for the induction and maintenance of adaptive immunity in a number of situations. Here, we examined STING expression by IHC, using a genetically validated assay, and show that the majority of atrophic prostatic luminal epithelial cells in all PIA lesions (N=20) consistently express high levels of STING, which is present in normal prostatic basal epithelial cells yet absent in normal-appearing prostatic luminal cells. In primary prostatic adenocarcinomas (> 260 tumors), STING protein was either completely negative , or extremely low in ~ 95% of cases. That STING in PIA is activate is supported by single cell RNA sequencing (scRNAseq) data from human prostatectomy tissues that show cells with the phenotype of intermediate luminal cells, indicative of PIA, express a number of interferon response genes that are not expressed in normal luminal cells (PMID 37905029). To our knowledge this is the first systematic report of STING protein staining in human prostate cancer and benign tissues. We postulate that upregulation of STING in human intermediate cells in PIA represents an inflammatory response that can drive interferon stimulated gene expression and inflammation in PIA. This innate immune response is absent, potentially by gene silencing, in prostatic adenocarcinoma, which may help neoplastic cells evade the immune response and could help contribute to the long standing finding that most prostate cancers are immunologically cold. Citation Format: Thomas M. Steele, Alan K. Meeker, Mindy K. Graham, Rulin Wang, Carolina Gomes-Alexandre, William G. Nelson, Fernanda Caramella Pereiria, Sushant Kachhap, Srinivasan Yegnasubramanian, Angelo Michael De Marzo. STING protein expression is increased in intermediate luminal cells in proliferative inflammatory atrophy of the prostate (PIA) and is absent in the majority of primary prostatic adenocarcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB324.

Topical imiquimod treatment of residual or recurrent cervical intraepithelial neoplasia lesions (TOPIC-2): A randomised controlled trial.

To investigate the efficacy of imiquimod in women with residual or recurrent cervical intraepithelial neoplasia (rrCIN), compared with large loop excision of the transformation zone (LLETZ). Randomised controlled non-inferiority trial. One academic and one regional hospital in the Netherlands. Thirty-five women with rrCIN were included in the study between May 2016 and May 2021. Women were randomised to receive treatment with 5% imiquimod cream (12.5 mg) intravaginally (three times a week for a duration of 16 weeks) or a LLETZ procedure (standard treatment). The primary outcome was reduction to normal cytology at 6 months after starting treatment. Secondary outcomes were clearance of high-risk human papilloma virus (hr-HPV) in both groups and reduction to ≤CIN1 in the imiquimod group. Side effects were monitored. Treatment success was 33% (6/18) in the imiquimod group versus 100% (16/16) in the LLETZ group (P < 0.001), whereas HPV clearance was 22% (4/18) in the imiquimod group versus 88% (14/16) in the LLETZ group (P < 0.001). After the randomisation of 35 women, the futility of treatment with imiquimod was proven and the trial was prematurely finished. In the follow-up period, three patients remained without additional treatment, whereas all other patients underwent LLETZ, conisation or hysterectomy. In the LLETZ group none of the patients received additional treatment during 2 years of follow-up. This is the first randomised controlled trial to show that topical imiquimod has a significantly lower success rate in terms of reduction to normal cytology and hr-HPV clearance, compared with LLETZ, in women with rrCIN. Additionally, imiquimod has numerous side effects and after using imiquimod most women with rrCIN still required additional surgical treatment.

Abstract 5325: Cytokine CCL9, new downstream target of oncogene Kras to modulate acinar-to-ductal metaplasia during pancreatic cancer initiation

Abstract Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal types of cancer with a current five-year survival rate at 12% in the US, and is dominated by oncogenic Kras mutations. Accumulating evidence showed that PDAC can be originated from acinar-to-ductal metaplasia (ADM). In this process, pancreatic acini harboring oncogenic Kras mutations are transdifferentiated to a duct-like phenotype that further progresses to become pancreatic intraepithelial neoplasia (PanIN) lesions, giving rise to PDAC. Although ADM formation is commonly observed in KrasG12D transgenic mouse models of PDAC, the exact mechanisms of how oncogenic KrasG12D modulates this process remain unclear. Here, we unveil a new downstream target of oncogenic Kras, cytokine CCL9 during ADM formation. Either exogenously treating primary pancreatic acini with recombinant CCL9 or ectopically expressing CCL9 in primary pancreatic acini resulted in ADM formation in a 3D organoid culture system. Furthermore, knockdown of CCL9 in KrasG12D-expressed acini reduced KrasG12D-induced ADM. Higher levels of reactive oxygen species (ROS) were detected in the KrasG12D-expressed pancreatic acini, and knockdown of CCL9 in these acini decreased ROS levels. Overexpression of CCL9 in primary pancreatic acini elevated cellular ROS of pancreatic acinar cells. Furthermore, depletion of ROS in 3D organoid culture of CCL9-expressed acini reduced CCL9-induced ADM formation. In transgenic mice of p48cre:KrasG12D, blockade of CCL9 through its specific neutralizing antibody attenuated ADM formation as well as PanIN structures. Altogether, our results indicated oncogenic KrasG12D initiates PDAC through a new downstream target molecule, CCL9, to enhance ROS levels of pancreatic acinar cells. Citation Format: Geou-Yarh (Stancy) Liou, Justin K. Messex, Crystal J. Byrd. Cytokine CCL9, new downstream target of oncogene Kras to modulate acinar-to-ductal metaplasia during pancreatic cancer initiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5325.

Abstract 3447: Potential for reduction in invasive cancer incidence by interception of preneoplastic lesions

Abstract Background: Preneoplastic lesions (PNL) are morphologically identifiable lesions lack the ability to invade surrounding tissue but can progress to invasive cancer. Thus, PNL present an opportunity to identify individuals at elevated cancer risk and identify opportunities for early detection and interception of cancer. Objective: We used published data sources to estimate the impact of intercepting progression of PNL to invasive cancer across multiple anatomic sites. We estimated the hypothetical number of cancers that could be avoided in the United States population if interception was successful in preventing a percentage of individuals with PNL from progressing to invasive cancer. Methods: Estimates of prevalence, latency, and probability of percent progression of PNL to invasive cancer were obtained from literature. We considered PNL associated with invasive tumors of the breast, bladder, cervix, colorectum, esophagus, liver, lung, oropharynx, pancreas, prostate, skin (melanocytes), skin (squamous cells), and stomach as well as precursors to leukemia and myeloma. We modeled a cohort of 100,000 individuals from which we estimated the number of cases that could be avoided if interception of a PNL before it progressed to an invasive cancer case was successful. Results were stratified for race and sex; positive- and negative-predictive values were calculated. Results: For PNL with a higher prevalence and percent progression per year, there were more invasive cancers that could be avoided if interception was successful. Detection of PNL associated with pancreas, stomach, oral, and prostate cancers had the greatest potential to avoid invasive cancers in the US population. For example, if 95% of pancreatic intraepithelial neoplasia lesions were detected, 15% progressed to pancreas cancer, and assuming a 43% prevalence over a 5-year latency period, 1,226 potential invasive pancreatic cancer cases could be avoided per year. Similarly, 998 potential invasive stomach cancer cases could be avoided earlier if 95% of gastric intestinal metaplasia lesions were detected assuming 42% progressed to invasive cancer with a 19% prevalence and a 4-year latency period. Additionally, if we could intercept 5% to 95% of all PNL from becoming invasive cancer, we would avoid between 176 to 8,515 number of invasive cases being diagnosed. When stratified by race, symptomatic multiple myeloma can be avoided in individuals with monoclonal gammopathies of undetermined significance if this condition is detected earlier among Black individuals. Conclusion: By detecting PNL and preventing them from advancing to invasive cancer, we can avoid substantial mortality as well as the costs and morbidities associated with treating more advanced cancers. Citation Format: Pranoti Pradhan, Irene Ghobrial, Catherine Marinac, Betsy O’Donnell, Sapna Syngal, Timothy Rebbeck. Potential for reduction in invasive cancer incidence by interception of preneoplastic lesions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3447.

Exploring the potential prompting role of cervical human papilloma virus detection in vulvar lesions: a cross-sectional study in China.

The etiology and clinical presentation of vulvar carcinomas, especially vulvar lesions, are not fully understood. Because the vulva and cervix are anatomically connected, human papillomavirus (HPV) is the main cause of cervical lesions. Thus, this study explored the potential characteristics and effects of specific HPV infection types across vulvar lesions and concurrent cervical lesions. This retrospective, cross-sectional study analyzed patients with cervical HPV or cytological results and concurrent vulvar biopsy who were seen in our hospital colposcopy clinic in Shanxi Province, China, between 2013 and 2023. Data on age, menopause status, vulvar manifestations, and cytology and HPV infection testing results were collected. Attributable fractions and multinominal logistic models were used to evaluate HPV genotyping and clinical characteristics across vulvar lesions. Among the 1,027 participants, 83 (8.1%) had vulvar intraepithelial neoplasia (VIN) of high grade or worse (VIN2+), and 127 (12.4%) had non-neoplastic epithelial disorders of the vulva (NNEDV). A total of 175 patients had either VIN2+ or cervical intraepithelial neoplasia (CIN) lesions of grade 2 or worse (CIN2+). The most common HPV genotypes for VIN2+ or concurrent VIN2+/CIN2+ were HPV16, HPV52, and HPV58, although attributable fractions differed among lesions. Patients with normal cytological or histopathological result were more likely to have NNEDV detected, while abnormal cervical diagnosis was associated with higher detection of VIN2+. Multinominal logistic modeling showed that age and HPV16 infection were risk factors for VIN2+ or concurrent VIN2+/CIN2+; however, only vulvar presentation with depigmentation was a risk factor for NNEDV. Among patients with low-grade CIN1/VIN1, compared with those who were HPV16 negative, those who were HPV16 positive were at 6.63-fold higher risk of VIN2+/CIN2+ [95% confidence interval (CI): 3.32, 13.21]. Vulvar depigmentation was also associated with increased risk of NNEDV (odds ratio: 9.98; 95% CI: 3.02, 33.04). Chinese women may be at specific, high risk for HPV infection types associated with VIN or CIN. The use of cervical cell HPV detection along with vulvar presentation during cervical cancer screening may also contribute to vulvar lesion detection.

Abstract B113: Characterization of the epigenomic landscape of the human pancreas and early pancreatic neoplastic lesions

Abstract Study of early pancreas neoplasia in humans had previously been limited by lack of available tissue. Recent work by our group on deceased donor pancreata identified pancreatic intraepithelial neoplasia (PanIN) lesions in non-diseased organs and defined a transcriptomic signature for the microenvironment of these pre-cancerous lesions. Prevalence of PanINs in healthy human tissue was higher than expected and established a novel model to expand understanding of the complex biology of these precursor lesions. Epigenetic changes in chromatin structure and the subsequent effects on gene expression are essential processes in neoplasia that have yet to be described at the single cell level in the human pancreas. We hypothesized that characterization of the epigenomic landscape in donor pancreata would identify differences in chromatin structure and gene expression between healthy human tissue and PanINs, thereby expanding prior knowledge and identifying novel targets of study in the biology of early neoplasia in the pancreas. Single nuclei from five donor pancreata (six total samples including one replicate sample from a single donor pancreas) were isolated for transposase-accessible chromatin preparation followed by high-throughput sequencing (ATAC-seq) using the 10x Genomics platform. Data analysis was conducted using CellRanger, Seurat, Signac, and AUCell to identify and label chromatin peaks in individual nuclei corresponding to acinar, ductal, and PanIN origin according to gene signatures identified by scRNA on matched pancreata (Carpenter et al 2023, Cancer Discovery). Using this method, cells corresponding to these gene signatures, including PanIN-like cells, were indeed identified in preliminary analysis. Differentially accessible regions of the genome were then analyzed for motif enrichment between these cell types. Motifs enriched in cells identified as PanINs compared to either ductal or acinar cells included several transcription factors implicated in tumorigenesis, including members of the AP-1/Fos/Jun family and multiple zinc-finger protein family members, such as the KLF. These results are similar to previously demonstrated essential factors in pancreatic neoplasia. Differentially expressed motifs between ductal and acinar cells included factors associated with differentiation, including Myod1, Ptf1a, and the TCF family of transcription factors. Together, this study demonstrates that PanINs can be identified in human pancreas tissue by ATAC-seq and that differentially expressed motifs can be identified this model, providing opportunity for further integration with transcriptomic information to elucidate detailed understanding of early neoplasia in the pancreas. Citation Format: Jamie N. Mills, Joyce Thompson, Jacqueline Morales, Padma Kadiyala, Ahmed M. Elhossiny, Howard Crawford, Eileen Carpenter, Marina Pasca di Magliano, Filip Bednar, Simone Benitz. Characterization of the epigenomic landscape of the human pancreas and early pancreatic neoplastic lesions [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B113.

PHOTODYNAMIC THERAPY FOR CERVICAL CANCER: A LITERATURE REVIEW

Relevance: Cervical cancer is a major public health problem worldwide, with human papillomavirus infection playing a vital role as a risk factor. Photodynamic therapy is a minimally invasive treatment for HPV-related cervical lesions that uses photosensitizers and light to selectively destroy abnormal cells.&#x0D; The study aimed to comprehensively review the different types of molecules used in PDT to reduce the morbidity and mortality associated with cervical cancer.&#x0D; Methods: A comprehensive search was conducted for all relevant articles investigating the efficacy and safety of PDT in the treatment of HPV-associated cervical cancer. PICO scores were determined for the review, and a literature search of the PubMed database was performed. An examination of the PubMed online database using keyword combinations identified 71 studies conducted between 2013 and 2023 that investigated using PDT to treat RSM cells.&#x0D; This article reviews ongoing clinical trials examining the efficacy of PDT in treating low-grade squamous cell intraepithelial neoplasia and high-grade squamous cell intraepithelial lesions, as well as preclinical approaches using different molecules for PDT in cervical cancer&#x0D; Results: Potential molecules for PDT are described, their advantages and disadvantages evaluated, and solutions to improve their compatibility with antitumor treatment are proposed. Our review shows that PDT is a promising therapeutic approach for diagnosing and treating HPV-related cervical lesions. At the same time, we observe that using different classes of dyes enhances the anticancer effects of PDT&#x0D; Conclusion: Fullerene and ALA-PDT are potential leaders for more intensive use in PDT, which will further help reduce the global incidence and mortality from cervical cancer. However, further studies are needed to evaluate its long-term efficacy and safety.

A new histopathological phenomenon: Pancreatic islet cell loss in the elderly population

BackgroundWe observed the phenomenon of pancreatic islet cell loss (ICL) in our previous histopathological study. Multiple studies have reported that a decrease in β-cells is correlated with diabetes or chronic pancreatitis. Few studies have reported ICL in a healthy population. MethodsThirty-three pancreatic tissue samples were obtained from cadavers (age: 65–104 years) who had never been diagnosed with any pancreatic diseases before death. The pancreatic body sections were used for an immunohistochemical study of pancreatic islet cells, and area calculations were performed using ImageJ to determine the degree of ICL and islet cell proportions. ResultsThe proportion of β-cells showed a downward trend as the degree of ICL increased (r=−0.414, P = 0.011), and the proportion of women with severe ICL was significantly higher than that of men with severe ICL (P = 0.016). The probability of severe ICL decreased with age in the population over 70 years of age (P = 0.069, linear correlation). Severe ICL may be associated with higher pancreatic intraepithelial neoplasia lesions (P = 0.059). ConclusionThe phenomenon of ICL in the elderly population was mainly due to pancreatic β-cell reduction. It may be one of the direct causes of age-related diabetes.

Relative Telomere Length in Cervical Exfoliated Cells among Women with High-Risk Human Papillomavirus

Introduction: This study investigates and compares the relative telomere length (RTL) outcome of high-risk (hr) human papillomavirus (HPV)-infected normal, low-grade squamous intraepithelial lesion (LSIL), and high-grade squamous intraepithelial lesion (HSIL) cervical samples to HPV-free normal cervical samples. Methods: This study used archived cervical samples and obtained cytology and histology data. HPV genotyping was conducted using Sanger sequencing, and RTL was performed using real-time quantitative polymerase chain reaction. Results: This study investigated 287 cervical samples, including 100 normal and hr-HPV-negative samples from the control group, 44 normal and hr-HPV-infected samples, and 143 SIL and hr-HPV-infected samples. The RTL in hr-HPV-infected samples, including the SIL and normal sample groups, was significantly longer than that in the control group. RTL in HSIL (5.13 ± 3.22) and LSIL (2.86 ± 2.81) was significantly different (p < 0.001). The RTL of cervical intraepithelial neoplasia (CIN1) lesion (3.53 ± 2.53) differed significantly (p < 0.001) when compared to CIN2 and CIN3 lesions combined (12.04 ± 10.51). The risk of developing cervical cancer was associated with RTL and decreased with RTL. Conclusion: This study revealed the strong potential of the RTL test in identifying women at risk of developing cervical cancer.

Validation of an on-chip p16ink4a/Ki-67 dual immunostaining cervical cytology system using microfluidic device technology

More specific screening systems for cervical cancer may become necessary as the human papillomavirus (HPV) vaccine becomes more widespread. Although p16/Ki-67 dual-staining cytology has several advantages, it requires advanced diagnostic skills. Here, we developed an automated on-chip immunostaining method using a microfluidic device. An electroactive microwell array (EMA) microfluidic device with patterned thin-film electrodes at the bottom of each microwell was used for single-cell capture by dielectrophoresis. Immunostaining and dual staining for p16/Ki-67 were performed on diagnosed liquid cytology samples using the EMA device. The numbers of p16/Ki-67 dual-stained cells captured by the EMA device were determined and compared among the cervical intraepithelial neoplasia (CIN) lesion samples. Seven normal, fifteen CIN grade 3, and seven CIN grade 2 samples were examined. The percentage of dual-positive cells was 18.6% in the CIN grade 2 samples and 23.6% in the CIN grade 3 samples. The percentages of dual-positive staining increased significantly as the severity of the cervical lesions increased. p16/Ki67 dual immunostaining using the EMA device is as sensitive as the conventional method of confirming the histopathological diagnosis of cervical samples. This system enables a quantified parallel analysis at the individual cell level.

Foamy Gland Adenocarcinoma with High-Grade Prostatic Intraepithelial Neoplasia (HGPIN) Lesion on TRUS Core Needle Biopsy: A Rare Pathological Variant

Foamy gland carcinoma is a variant of prostatic acinar adenocarcinoma characterized by abundant, foamy cytoplasm, frequently showing small, pyknotic nuclei. Foamy gland carcinoma may be deceptively benign appearing and missed on needle biopsy. Most foamy gland carcinomas are Gleason score 6 or 7, although Gleason score 8-10 carcinomas have been reported. Its occurrence is extremely rare. Prognosis depends on the Gleason score and the presence or absence of perineural or extra prostatic extension. Treatment modalities of foamy gland adenocarcinoma are a similar to that of usual acinar adenocarcinoma. We report an extremely rare case of foamy gland adenocarcinoma of the prostate associated with high-grade prostatic intraepithelial neoplasia lesion on TRUS core needle biopsy in a 78-year-old man presented with history of lower urinary tract symptoms (LUTS) during the last 6 months. Digital rectal examination: the prostate was enlarged, firm in consistency, non-tender, but with a nodule on its left base. Serum prostate-specific antigen (PSA) was 19 ng/mL. Prostate RMI showed a PIRADS 4 lesion on the left base of the prostate. TRUS-guided core biopsy was performed. Pathology report of the prostate cores found foamy gland adenocarcinoma; Gleason score 9 (4 +5) with presence of high-grade PIN lesion. The Bone scan and chest/abdomen CT scan were unremarkable. There were a significant relief of his LUTS under alpha-blockers. After discussion of all the management options, the patient chose a simple surveillance with a close follow-up for LUTS and PSA every 3months and thereafter acting accordingly.

The Fate of Cervical Dysplastic Lesions during Pregnancy and the Impact of the Delivery Mode: A Review.

Cervical dysplasia, also referred to as cervical intraepithelial neoplasia (CIN) or squamous intraepithelial lesion (SIL), is the precursor lesion of cervical carcinoma. Therefore, its diagnosis is vital for early detection and inhibiting the development of cervical carcinogenesis. Human papillomavirus (HPV) is the most common aetiology of cervical cancer and this infection mainly affects young women of childbearing age, thus affecting pregnant women as well. It is essential to know how CIN progresses in pregnant patients because the management of pregnant and non-pregnant patients is different (considering the safety of both mother and child in pregnancy). This review intends to highlight the studies which have assessed the rates of progression of CIN diagnosed in pregnancy throughout the antenatal period and the impact of the mode of delivery on CIN outcomes. We searched PubMed/MEDLINE and Google Scholar databases for relevant articles. Many studies indicate that the rate of progression of these lesions is very slow during the tenure of pregnancy; many also report postpartum regression of these lesions. Thus, in most of these patients, management can be safely implemented in the postpartum period while just keeping them under observation in the antenatal period. However, patients with high-grade CIN have a higher chance of developing invasive cancer and, therefore, require careful monitoring. There is a dispute regarding the role of the mode of delivery in determining the fate of cervical dysplasia. While some studies supported vaginal births over caesarean sections, others did not find any difference between the two in defining the outcome of the dysplastic lesions.

Pivotal Clinical Study to Evaluate the Efficacy and Safety of Assistive Artificial Intelligence-Based Software for Cervical Cancer Diagnosis.

Colposcopy is the gold standard diagnostic tool for identifying cervical lesions. However, the accuracy of colposcopies depends on the proficiency of the colposcopist. Machine learning algorithms using an artificial intelligence (AI) system can quickly process large amounts of data and have been successfully applied in several clinical situations. This study evaluated the feasibility of an AI system as an assistive tool for diagnosing high-grade cervical intraepithelial neoplasia lesions compared to the human interpretation of cervical images. This two-centered, crossover, double-blind, randomized controlled trial included 886 randomly selected images. Four colposcopists (two proficient and two inexperienced) independently evaluated cervical images, once with and the other time without the aid of the Cerviray AI® system (AIDOT, Seoul, Republic of Korea). The AI aid demonstrated improved areas under the curve on the localization receiver-operating characteristic curve compared with the colposcopy impressions of colposcopists (difference 0.12, 95% confidence interval, 0.10-0.14, p < 0.001). Sensitivity and specificity also improved when using the AI system (89.18% vs. 71.33%; p < 0.001, 96.68% vs. 92.16%; p < 0.001, respectively). Additionally, the classification accuracy rate improved with the aid of AI (86.40% vs. 75.45%; p < 0.001). Overall, the AI system could be used as an assistive diagnostic tool for both proficient and inexperienced colposcopists in cervical cancer screenings to estimate the impression and location of pathologic lesions. Further utilization of this system could help inexperienced colposcopists confirm where to perform a biopsy to diagnose high-grade lesions.

Abstract A028: Dissecting the role of cellular senescence in prostate cancer initiation and immune suppression

Abstract Prostate cancer is the most common malignancy and the second leading cause of cancer-related death in American men. Although many disease cases remain latent for decades, others progress to malignant prostate cancer with high rates of mortality. The molecular underpinnings driving malignancy are still poorly understood, and there is an urgent need to identify mechanisms and informative biomarkers of disease progression. Using the Pb-Cre+; Ptenfl/fl (CP) mouse model, we and others found that Pten genomic loss, a common occurrence in human prostate cancer, leads to induction of cellular senescence, senescence-associated inflammation, and immune suppression during tumor onset. Similarly, senescence markers have also been identified in benign and early prostate cancer lesions in humans. This suggests that senescence may play a yet uncharacterized functional role in prostate tumorigenesis. Cellular senescence is a damage-induced pathway leading to durable growth arrest and has long been considered a potent tumor suppressive mechanism that limits the ability of pre-malignant cells to proliferate and develop into malignant tumors. However, recent findings suggest that the accumulation of senescent cells can paradoxically lead to chronic inflammation that fosters an environment for eventual tumor development and progression through the induction of growth and inflammatory factors collectively known as the senescence-associated secretory phenotype (SASP). Thus, it remains unclear whether senescence or specific senescent cell populations block or rather promote prostate tumor development. Leveraging the CP prostate cancer mouse model and genetic tools to track senescent cells, we observed that markers of senescence (SA-β-gal, p16, Bcl2) and the inflammatory SASP (NF-𝜅B, IL-1b, CXCL1, CSF-1) were not only present in early prostate intraepithelial neoplasia (PIN) lesions but remained elevated following adenocarcinoma development and progression, and could be found in both epithelial cells as well as other immune and stromal populations. Expression of the canonical senescence marker p16 also correlated with disease progression in human prostate cancer samples. Moreover, senescent cell accumulation was associated with a reduction in cytotoxic lymphocytes and an influx of suppressive myeloid populations that we and others have previously shown contribute to prostate cancer progression. Senolytic approaches to eliminate these senescent cell populations reduced progression to adenocarcinoma and remodeled the immune system to reverse myeloid-mediated immune suppression and activate cytotoxic lymphocyte immunity. Together, our preliminary results suggest that senescence might contribute to prostate cancer initiation and progression, and that removal of senescent cells could delay tumor onset and reverse prostate cancer immune suppression. Citation Format: Lin Zhou, Jarin Snyder, Katherine C. Murphy, Kelly D. DeMarco, Sachliv Chana, Karl Simin, Zhong Jiang, Marcus Ruscetti. Dissecting the role of cellular senescence in prostate cancer initiation and immune suppression [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr A028.

Distribution of cervical intraepithelial neoplasia is closely associated with HPV status and uterine position.

Although cervical intraepithelial neoplasia (CIN) lesions are considered to be not randomly distributed across the cervix, but predominantly in the anterior wall, the clinicopathological etiology remains unknown. Herein, we aimed to elucidate the relationship between quantitatively measured area of CIN2/3 and cervical cancer associated factors by retrospective cohort study. We analyzed 235 consecutive therapeutic conization specimens dissected as a single intact section to determine CIN2/3 area and its correlation with both clinical risk factors including human papillomavirus (HPV) status (single or multiple infection) and uterine position defined by transvaginal ultrasound. Cervical wall was classified into three groups: anterior: (11, 12, 1, and 2 o'clock), posterior (5, 6, 7, and 8 o'clock) and lateral (3, 4, 9, and 10 o'clock). Multiple regression revealed that younger age and HPV16 status were significantly correlated with CIN2/3 area (p = 0.0224 and p = 0.0075, respectively). The Jonckheere-Terpstra test showed a significant trend: CIN2/3 area was highest in the single HPV16 group, followed by the multiple HPV16 group and the non-HPV16 group (p < 0.0001). CIN2/3 area in the anterior wall was statistically significantly larger than the posterior and lateral wall (p = 0.0059 and p = 0.0107, respectively). CIN2/3 area in the anterior wall was significantly greater with anteversion-anteflexion than retroversion-retroflexion (p = 0.0485), whereas CIN2/3 area in the posterior wall was significantly larger with retroversion-retroflexion than anteversion-anteflexion (p = 0.0394). In conclusion, the topographical distribution of CIN2/3 area is closely associated with patient age, high-risk HPV status, especially single HPV16 infection and uterine position.

Major hurdles to the use of tyrosine kinase inhibitors in clinical prevention/interception studies: Do preclinical studies with EGFR inhibitors suggest approaches to overcome some of the limitations.

There are major hurdles to the use of tyrosine kinase inhibitors (TKIs) and any other agents with significant toxicities (which means practically the preponderance of potential effective agents) in the context of prevention/anti-progression (interception) studies. We will discuss epidermal growth factor receptor (EGFR) inhibitors as examples, both in a primary prevention setting, where agent(s) are administered to individuals with no cancer but who might be considered at higher risk due to a variety of factors, and in anti-progression/interception studies, where agent(s) are administered to persons with known preinvasive lesions (e.g., colon adenomas, lung nodules, ductal carcinoma in situ (DCIS), or pancreatic intraepithelial neoplasia (PanIN) lesions in the pancreas) in an attempt to reverse or inhibit progression of these lesions. Multiple potential hurdles will be examined, including: a) toxicity of agents, b) the likely range of subtypes of cancers affected by a given treatment (e.g., EGFR inhibitors against EGFR mutant lung adenocarcinomas), c) the availability of practical endpoints besides the blocking of cancer formation or pharmacokinetics related to the agents administered in a primary prevention study, and d) the interpretation of the regression or blockage of new preinvasive lesions in the anti-progression study. Such an anti-progression approach may help address some of the factors commented on regarding primary prevention (toxicity, potential target organ cancer subtypes) but still leaves major questions regarding interpretation of modulation of preinvasive endpoints when it may not be clear how frequently they progress to clinical cancer. Additionally, we address whether certain recent preclinical findings might be able to reduce the toxicities associated with these agents and perhaps even increase their potential efficacy. Antibodies and TKIs other than the EGFR inhibitors are not discussed because few if any had been tested as monotherapies in humans, making their efficacy harder to predict, and because a number have relatively rare but quite striking toxicities. Furthermore, most of the practical hurdles raised regarding the EGFR inhibitors are relevant to the other TKIs. Finally, we briefly discuss whether early detection employing blood or serum samples may allow identification of high-risk groups more amenable to agents with greater toxicity.

Abstract 637: Cytokine CCL9 mediates oncogenic Kras-induced pancreatic acinar-to-ductal metaplasia through ROS

Abstract Pancreatic ductal adenocarcinoma (PDAC) can be originated from acinar-to-ductal metaplasia (ADM). Pancreatic acini harboring oncogenic Kras mutations are transdifferentiated to a duct-like phenotype that further progresses to become pancreatic intraepithelial neoplasia (PanIN) lesions, giving rise to PDAC. Although ADM formation is frequently observed in KrasG12D transgenic mouse models of PDAC, the exact mechanisms of how oncogenic KrasG12D regulates this process remain an enigma. Herein we reveal a new downstream target of oncogenic Kras, cytokine CCL9 during ADM formation. Exogenously treating primary pancreatic acini with recombinant CCL9 in a 3D organoid culture system drives ADM formation. Furthermore, knockdown of CCL9 in KrasG12D-expressed acini reduced KrasG12D-induced ADM. Higher levels of reactive oxygen species were detected in the KrasG12D-expressed pancreatic acini, and knockdown of CCL9 in these acini dramatically decreased ROS levels. Depletion of ROS in 3D organoid culture of CCL9-expressed acini reduced CCL9-induced ADM formation. In transgenic mice of p48cre:KrasG12D, depletion of CCL9 through its specific neutralizing antibody attenuated ADM formation as well as PanIN structures. Hence, our results provided insight into how oncogenic Kras regulates pancreatic ADM through a new downstream target molecule, CCL9. Citation Format: Geou-Yarh Liou, Justin K. Messex, Kiyah L. Adams. Cytokine CCL9 mediates oncogenic Kras-induced pancreatic acinar-to-ductal metaplasia through ROS [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 637.

Abstract 1243: RON receptor signaling enhances prostate cancer metastasis in Hi-Myc mice

Abstract Prostate cancer (PCa) is the second leading cause of cancer mortality among men globally. Death from PCa is typically due to the lack of effective treatments for advanced forms of prostate cancers including castration-resistant prostate cancer (CRPC) and metastatic prostate cancers (mPCa). Distantly metastasized cancers drop the 5-year survival rate from 98% to 30%. Understanding the intrinsic and extrinsic mechanisms that lead to the development of these advanced prostate cancers could improve treatment options. The RON receptor tyrosine kinase is a cell surface receptor, which is activated by growth factor ligands (HGFL or Gas6), which binding promotes several cancer phenotypes in various cancers including prostate cancer. Our laboratory has shown that RON is overexpressed in 86% of primary and 100% of metastatic human prostate cancers compared to normal prostate tissue. Additionally, we have shown that RON overexpression in the prostates of mice can induce Prostatic intraepithelial neoplasia (PIN) lesions, adenocarcinoma, and increases several cancer phenotypes including proliferation, cell survival, metastasis, and increased recruitment of tumor-associated macrophages. However, we have yet to explore the effect of RON on prostate cancer metastasis. We hypothesize that RON expression enhances metastasis by promoting an increase in the amount of disseminating prostate cancer cells. To test this hypothesis, the Hi-Myc mouse model has been employed. The MYC gene is one of the most dysregulated genes in prostate cancers and Myc expression has been detected in the early stages of prostate cancer development, PIN lesion, and all later stages of prostate cancer. Additionally, the Hi-Myc model is clinically relevant as prostate tumor development in this model is similar to that in humans in regard to progression from PIN lesions to adenocarcinoma. From the Hi-Myc model, we have generated two additional genetic modifications within prostate epithelial cells: 1) Hi-Myc mice with prostate-specific RON overexpression (OE) and 2) Hi-Myc mice with prostate-specific RON loss (ΔEpi). In this study we report that a lack of prostate RON expression diminishes adenocarcinoma development and metastasis within the Hi-Myc mouse model. Moreover, RON overexpression promotes the development of metastatic adenocarcinoma that was not observed in the prostates from Hi-Myc control mice. These studies suggest that RON expression supports the advancement of primary tumors to metastatic prostate cancer within the Hi-Myc mouse models. Therefore, future directions include determining the signaling pathways driven by RON that promotes this metastatic phenotype in both castration-sensitive and resistant tumors. Citation Format: Angelle D. Jones, Carissa Lester, Susan E. Waltz. RON receptor signaling enhances prostate cancer metastasis in Hi-Myc mice [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 1243.

Abstract 4230: Transcriptomic evaluation of exercise-induced suppression of prostate cancer aggressiveness

Abstract Previous studies from our laboratory have showed that aerobic exercise significantly reduced the number of aggressive poorly differentiated tumors in the transgenic adenocarcinoma of the mouse prostate (TRAMP) model. Despite these encouraging data the underlying mechanism of how exercise reduces tumor aggressiveness remains undefined. We aimed to fill this scientific gap by utilizing a transcriptomics approach to identify potential mechanisms by which aerobic exercise suppresses prostate tumor aggressiveness. Methods: Twelve TRAMP mice, 8-10 weeks of age, were equally randomized to exercise or control group. Mice in the exercise group were singularly housed in cages with running wheels for 12 weeks. Mice in the control group maintained normal group housing and activity conditions for 12 weeks. At euthanasia, prostate tumors were excised, weighed and processed for immunohistochemistry and transcriptome analysis. Two independent pathologists, blinded to the interventions, performed histological analysis of the genitourinary mass. Outputs of sequencing data were assessed for quality and accuracy. Counts for all known mRNA, differential expression, and heatmap were prepared. Differential expression was filtered to identify genes that had a ≥2-fold change with an adjusted p&amp;lt;0.05. Gene ontology and pathway analyses was performed to reveal selective pathways activated. Results: No significant difference in genitourinary mass, body mass or tumor free body mass was found between groups. Pathology revealed majority of the tissue from the control group exhibited moderate to poorly differentiated tumors (3/6). On the other hand, none of the animals in the exercise intervention group showed such pathology. Four out of five showed well differentiated tumors including prostatic intraepithelial neoplasia (PIN) lesions in one animal. Transcriptomic analysis coupled with gene set enrichment identified pathways associated with triglyceride catabolic process, lipid homeostasis, lipid metabolic process, triglyceride metabolic process to be most impacted. Differentially expressed genes of interest include haptoglobin (HP) and hormone sensitive lipoprotein lipase (Lipe) were significantly lower in the exercise group. Conclusion: Our preliminary findings provide novel evidence suggesting that exercise suppresses prostate tumor aggressiveness, in part, through transcriptomic modulation and altered cellular pathways associated with intratumoral energy metabolism. This project was supported by the National Center Institute designated Mays Cancer Center at UT Health San Antonio. Citation Format: Darpan I. Patel, Paul Rivas, Yidong Chen, Zhao Lai, Robert L. Reddick, Yuji Ikeno, Rita Ghosh, A. Pratap Kumar. Transcriptomic evaluation of exercise-induced suppression of prostate cancer aggressiveness. [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 4230.