Primary Screening Research Articles

The current state of the problem of cervical screening

the article summarizes the issue of prevention of cervical cancer - the most common disease associated with the human papillomavirus (HPV). The main goal of this study is to generalize the etiology of cervical cancer, to note unique opportunities for its prevention, as well as to develop screening tests for HPV as one of the most effective strategies. Explore how current HPV diagnostics detect E6/E7 DNA or mRNA in cervical/vaginal specimens using molecular technologies. To find out whether HPV screening tests are more sensitive than cytology or visual examination with acetic acid as the primary screening method, and even more clinically valuable for detecting subtle cytological changes as a hybrid test. Through a detailed study of literature sources, their systematization and approaches to solving the problem, it was found that technical and laboratory capabilities are very limited in underserved places, so women have to travel long distances and make some efforts to prevent and treat cervical cancer. The relevance of the study of this pathology lies in the fact that cervical cancer is a serious public health problem and is the fourth most common cancer in women worldwide, accounting for about 604,000 new cases every year. And the practical implementation of an HPV screening program can face many problems, and measures must be taken to overcome these problems without compromising the detection of the disease. These measures could include reducing the frequency of screening using the WHO global strategy to offer HPV screening tests at ages 35 and 45, implementing high-throughput testing technology, and improving access to vaginal HPV screening tests for women in remote areas. Improving informational education for those who do not want to undergo a gynecological examination. Another important strategy is to implement a see-and-treat approach using a healthcare delivery platform that requires limited laboratory technician skills. In addition, the development and large-scale implementation of more specific HPV testing technologies that are much cheaper and easier to use in a non-laboratory setting than currently available options should be a priority for underserved facilities. At the same time, there is a need to develop and initiate implementation of an affordable and readily available intermediate or secondary test with optimal specificity to triage or separate clinically insignificant HPV infections that do not require colposcopy. The results of this study may be of interest to primary care physicians, gynecologists, oncologists, and therapists.

Breast Tumour Segmentation Using Advanced UNet with Saliency, Channel, and Spatial Attention Models

Cancers are getting pretty common these days and in that the second most common cancer in the world after lung cancer is breast cancer. The primary screening techniques for early diagnosis of cancerous nodules in women breast are Ultrasound, Mammography, and MRI analysis. However, accurately identifying and outlining the boundaries of tumor regions remains challenging due to their unpredictable shapes, random variations, and blurry outlines. Currently, the CAD based image analysis systems have gained huge attention in biomedical image processing systems along with Machine Learning (ML) and Deep Learning (DL) methods. The conventional machine learning methods suffer from accuracy related issues therefore deep learning-based schemes have been adopted. Specifically, UNet based approach is widely employed for biomedical image segmentation. This research focuses on development of a UNet based deep learning architecture for breast cancer image analysis and segmentation. The proposed model improves the segmentation accuracy by adding saliency model, channel and spatial attention model to consider the low- and high-level features. These modules help to achieve the clear boundary during segmentation. The performance of proposed model is evaluated by using publicly available mammogram dataset known as DDSM dataset.

Milk-based culture of Penicillium camemberti and its component oleamide affect cognitive function in healthy elderly Japanese individuals: a multi-arm randomized, double-blind, placebo-controlled study.

Dairy products and fermented foods have a reported association with maintained cognitive function. Camembert cheese, a dairy product fermented by the white mold Penicillium camemberti, has also been shown to enhance cognitive function in vivo. Oleamide, derived from the fermentation of the white mold, is a candidate for an active component, and expected to improve both cognitive function and sleep conditions. Thus, this study investigated whether the milk-based culture of white mold (MCW), and oleamide, could improve cognitive function and sleep state clinically. A multi-arm randomized, double-blind, placebo-controlled trial was conducted in Tokyo, Japan. 60 healthy Japanese individuals aged 50-75 who were aware of their cognitive decline were randomly and equally divided into three groups of 20 participants using computer-generated random numbers. Participants took either MCW (equivalent to 60 μg/day of oleamide), 60 μg/day of oleamide, or placebo capsules for 12 weeks. Serum BDNF, cognitive function by Cognitrax as primary and MCI Screen as secondary outcome, and sleep status using the Japanese version of the Pittsburgh Sleep Quality Index (PSQI-J) were assessed before and after intervention. The participants, outcome assessors and analysts, and research assistants were blinded to the group assignment. Of the 60 participants, 58 completed the study and were analyzed. No adverse events related to test foods were observed. The placebo group showed a negative rate of change in serum BDNF (-10.5% ± 19.7%), whereas the MCW and oleamide groups showed positive changes (2.0% ± 27.1% and 1.3% ± 13.5%, respectively). Cognitrax scores increased after 12 weeks in all groups. Conversely, the MPI score of the MCI Screen demonstrated a significant improvement in the MCW and oleamide groups compared to the placebo group (p = 0.013 and p < 0.001, respectively). The subscales, immediate free recall and delayed free recall, also significantly increased in them compared to the placebo group. Although PSQI-J revealed no significant differences among groups, the MCW and oleamide groups showed significant improvement after intervention in overall score, subjective sleep quality, and sleep latency. Our results suggest that MCW and its component, oleamide, are safe and contribute to maintaining cognitive functions, particularly short-term and working memory, and improving sleep state. Clinical trial registration: https://center6.umin.ac.jp/cgi-open-bin/ctr/ctr_view.cgi?recptno=R000054792, identifier UMIN-CTR UMIN000048084.

Urinary Incontinence in Primary Care-The Gap Between Recommendations and Real World.

Routine screening for urinary incontinence (UI) by primary care providers (PCPs) is recommended. We aimed to describe the rate of incident UI diagnosed at annual PCP visits, the prevalence of UI in a large primary care population, and estimate the rate of screening for UI during primary care preventive and annual wellness visits. Secondary aims were to describe PCP knowledge and behavior as they relate to UI screening and diagnosis. The electronic health record was used to abstract the number of adult female patients seen by PCPs within a regional health system with a diagnosis of UI before our study period and with a new diagnosis over a 2-year period. Additional new diagnoses and screening practices were found on chart review of an additional 824 representative charts. Primary care providers within the health system were surveyed about their screening practices and knowledge about UI. There were 192,053 women primary care patients seen over 2 years. A total of 5.7% had a UI diagnosis preceding the study period and 3.4% had a UI diagnosis during the study period. A total of 42% of PCPs reported that they screen for UI at least half the time and none were completely satisfied with their ability to screen for UI. Sixteen percent of annual wellness visits had any documentation of screening for UI. In a large primary care population, screening for and detection of UI in women was low.

Primary Care Screening Recommendations for People Living With Human Immunodeficiency Virus

Historically, a diagnosis of human immunodeficiency virus meant early death. However, advances in therapy have lengthened the life expectancy of people living with human immunodeficiency virus (PLWH), and primary care providers often manage their care. Human immunodeficiency virus infection increases the risk of developing coronary artery disease, cardiac arrhythmias, diabetes mellitus, osteoporosis, neuropathy, kidney disease, and cancer at a younger age. Consequently, PLWH require a distinct set of screening recommendations. Primary care providers should be aware of these recommendations to prevent debilitating sequelae and premature death associated with chronic diseases in PLWH.

Abstract 5858: Systematic identification of pathways associated with antibody drug conjugate sensitivity in breast cancer

Abstract Antibody drug conjugates (ADCs) are revolutionizing precision therapeutics in oncology. Sacituzumab Govitecan (SG), combining a Topoisomerase I (TOP1) inhibitor payload (SN38) and the tumor-selective antigen TROP2-targeting antibody hRS7, has demonstrated efficacy in challenging cancers including triple negative breast cancer (TNBC). Despite its promising clinical efficacy, the intricacies of sensitivity and resistance mechanisms to SG remain elusive. To systematically interrogate ADC sensitivity mechanisms, we employed genome-wide CRISPR-Cas9 knockout screening to discover novel regulators influencing SG sensitivity in human TNBC cells. Interactome analysis of top SG sensitizing hits showed clustered pathways of interest, including DNA repair/replication, mTORC1/metabolism, ubiquitin/proteolysis and endosome trafficking/sorting. A top druggable hit sensitizing to SG was PARP1, encoding the poly (ADP-ribose) polymerase 1 that is a key enzyme required for single-strand break repair pathways and for DNA replication fork stability. Furthermore, the synergistic lethality between SG and PARP1 was confirmed in experiments combining SG with PARP inhibitor treatment of TNBC cell lines. To selectively identify SG/ADC-specific genes and pathways, we performed a secondary round of CRISPR knockout and activation screens using a custom library comprising top hits discovered in the primary screen, treating cells in parallel with either SG or cytotoxic payload SN38 alone. Pathway analysis of genes selectively modulating sensitivity to SG but not SN38 revealed novel mediators involved in TROP2 turnover and recycling. We then performed validation and mechanistic studies to elucidate how specific genetic permutations regulate antibody delivery and sensitivity to SG. In summary, employing a systematic CRISPR screening method, we uncovered numerous novel pathways associated with both resistance and sensitization, playing a role in ADC delivery and target protein turnover. Our current focus involves prioritizing druggable genes and pathways, laying the foundation for proof-of-concept studies that will combine ADCs with specific targeted therapeutics, aiming to enhance tumor-killing efficacy. Citation Format: Bogang Wu, Sheng Sun, Nayana Thimmiah, Aiko Nagayama, James T. Coates, Ting Liu, Win Thant, Elena Bitman, David Li, John Doench, Aditya Bardia, Leif W. Ellisen. Systematic identification of pathways associated with antibody drug conjugate sensitivity in breast cancer [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 5858.

Abstract 6345: In vitro characterization of a dual antagonistic anti-LILRB2/LILRB4 monoclonal antibody

Abstract Introduction: Myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment are a potential therapeutic target in immune checkpoint cancer therapy, but MDSC-targeted therapies have yet been shown to improve survival. The leukocyte immunoglobulin (Ig)-like receptor B2 is a member of the leukocyte Ig-like receptor (LILR) family and is predominantly expressed on the surface of cells of the myelomonocytic lineage (monocytes, macrophages and dendritic cells). LILRB2 is a negative regulator of myeloid cells and is an immune checkpoint receptor. We chose to generate mAbs against LILRB2 as a potential immunotherapeutic for cancer. Methods: PentaMice® strains (5 WT mouse strains comprising 9 MHC class II haplotypes to maximize Ab diversity) were immunized with human LILRB2. Lymphocytes were fused with a myeloma partner to generate lead monoclonal hybridomas. 24 purified anti-LILRB2 mAbs were generated and evaluated by arrayed surface plasma resonance (SPR, Carterra® LSA® instrument) to assign epitope bins. mAb humanization was performed in silico to maximize T20 “humanness” scores (algorithm developed by Curia). Humanized mAbs were transiently expressed using Curia’s TunaCHO™ platform and tested in in vitro MDSC assays. Autologous co-cultures of magnetic bead enriched-human blood-derived MDSC (HLA-DR-CD14+) model cells and T cells (CD3+) were stimulated with plate-bound anti-CD3/CD28 to induce T cell activation. Cytokine secretion was assessed 4 days later. Results: A total of 384 hybridoma supernatants were selected from the primary screen. 245 were confirmed by ELISA and 178 stained HEK293 cells expressing LILRB2 cells by flow cytometry (FACS). 24 hybridomas were further selected, cloned and sequenced. Five epitope bins were identified via Carterra® LSA® instrument. Two clones were chosen for humanization, designated 6-I11A and 4-O19A and were recombinantly expressed with the human IgG4 isotype. FACS analysis on HEK293 cells expressing either huLILRB2 or huLILRB4 revealed that clone hu4-O19A-IgG4 was specific for huLILRB2. Surprisingly, clone hu6-I11A-IgG4 stained both huLILRB2 and huLILRB4. FACS analysis with hu4-O19A-IgG4 on human MDSCs exhibited an EC50 of 11.9 nM and a KD of 1.5 nM in binding to recombinant huLILRB2 antigen, while hu6-I11A-IgG4 had an EC50 of 3.2 nM and a KD of 0.6 nM. In co-cultures with human MDSCs and T cells, both hu6-I11A-IgG4 and hu4-O19A-IgG4 reversed cytokine suppression by the MDSCs by increasing GM-CSF, IL-10, TNFα, and IL-6, while suppressing IL-13, and had little effect on IL-17A, MIP-3α, IL-4, IL-2 and IFNγ compared to treatment with the isotype control, with hu6-I11A-IgG4 having a greater effect as an antagonist than hu4-O19A-IgG4. Conclusions: We have identified a dual anti-LILRB2/LILRB4 antagonist. Targeting MDSCs with this antibody offers great promise for immunotherapy for cancer as well as its immense potential for synergy with PD-1 blockade. Citation Format: Bruce L. Daugherty, Xiaomei Ge, Christine L. Hsieh, Brian A. Zabel, Seth Lederman. In vitro characterization of a dual antagonistic anti-LILRB2/LILRB4 monoclonal antibody [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 6345.

Abstract 3449: Is AI-enhanced breast ultrasound ready for breast cancer screening in low-resource environments? A systematic review

Abstract Purpose. Screening mammography is unavailable in many low-resource areas. We ask if the state-of-the-art in artificial intelligence (AI)-enhanced breast ultrasound (BUS) is sufficiently accurate to be used for primary breast cancer screening in low-resource regions. Background. Since the 1980s, high-income countries have implemented mammographic screening programs, leading to breast cancer mortality reduction in screened women.1 Mammography is unavailable in many low-resource regions, such as the USAPI. Furthermore, travel difficulties and lack of radiologists hinder implementation. AI combined with portable BUS may address limitations of the high-income paradigm. In this systematic review, we ask if AI-enhanced BUS can detect/segment lesions (Objective 1) and classify lesions as cancerous (Objective 2). Methods. Two reviewers independently assessed articles from 1/1/2016 to 8/6/2023 from PubMed, Google Scholar, and citation searching. Studies which report on AI development and report performance on a patient-wise, held-out test set met the inclusion criteria. Studies were characterized by AI task and clinical application time. Dataset composition and performance were examined via narrative data synthesis. QUADAS-2 bias assessment was performed using criteria for each AI task. Success in (2) is defined by meeting minimum screening performance guidelines.2,3 Results. PubMed yielded 281 studies, Google Scholar yielded 225 studies, and a manual citation search yielded 41 studies. From 382 unique full texts evaluated, 52 articles met all inclusion criteria: 3 frame selection, 2 real-time detection, 2 combination, 14 segmentation-only, and 31 classification-only. Lesion segmentation-only models achieved a 90th percentile Dice similarity coefficient of 0.913 on generally small datasets. The best evidence for lesion cancer classification reported 0.976 area under the curve. All studies faced elevated bias and applicability concerns under QUADAS-2. Conclusion. Reported performance for (1) is insufficient to introduce AI-enhanced BUS for breast cancer screening. Evidence supporting AI-enhanced BUS for (2) is dependent on few studies relying on internal datasets, limiting generalizability. Geographically diverse clinical trials are needed to confirm and improve robustness of performance of AI-enhanced BUS for (1) and (2). References. 1. Marmot MG, et al. The benefits and harms of breast cancer screening: an independent review. British journal of cancer. 2013;108(11):2205-2240. 2. Lehman CD, et al. National Performance Benchmarks for Modern Screening Digital Mammography: Update from the Breast Cancer Surveillance Consortium. Radiology. 2017-04-01 2017;283(1):49-58. doi:10.1148/radiol.2016161174 3. Rosenberg RD, et al. Performance Benchmarks for Screening Mammography. Radiology. 2006-10-01 2006;241(1):55-66. doi:10.1148/radiol.2411051504 Citation Format: Arianna Bunnell, Dustin Valdez, Fredrik Strand, Yannik Glaser, Peter Sadowski, John A. Shepherd. Is AI-enhanced breast ultrasound ready for breast cancer screening in low-resource environments? A systematic review [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 3449.

SAEM systematic online academic resource (SOAR) review: Gastrointestinal illnesses.

Free open access medical education (FOAM) has become an essential tool for emergency medicine (EM) education and can be valuable to clinicians as a point-of-care resource. The development of the revised Medical Education Translational Resources Impact and Quality (rMETRIQ) tool provides a standardized means of quality assessment. Previous entries of the Society for Academic Emergency Medicine systematic online academic resource (SOAR) series have focused on renal, endocrine, and sickle cell disorders. In this iteration, we strive to identify, curate, and describe FOAM topics specific to acute gastrointestinal (GI) illnesses. We searched 389 keywords across 11 GI topics that were modified from the 2019 Model of the Clinical Practice of EM (EM Model) using the search engine Google FOAM and within the top 50 websites listed on Academic Life in Emergency Medicine's Social Media Index. The sites underwent preliminary screening to eliminate resources that were not relevant to EM or GI illnesses. Identified resources were evaluated with the rMETRIQ tool by five board-certified EM physicians who received rMETRIQ tool rater training. After duplicates of the initial 39,505 resources were eliminated, 8059 remained. Primary screening resulted in a final 1202 resources. The most common categories were large bowel (18%), small bowel (13%), stomach (11%), esophagus (11%), biliary (11%), and liver (10%). Many resources covered multiple topics and subtopics. The final mean intraclass correlation coefficient among the five physicians was 0.95 (95% CI 0.92-0.98) for rMETRIQ scoring. We identified 256 sites considered "high quality" with a rMETRIQ score of 16 or higher as designated in prior reviews. This iteration of the SOAR review resulted in the highest number of high-quality resources compared to other SOAR reviews, with 21% of resources thus far scoring ≥ 16. A final list of high-quality resources can guide trainees, educator recommendations, and FOAM authors.

Abstract 3094: Biophysics in kinase drug discovery programs: Spectral shift technology from fragment screening to mechanism of action

Abstract Using Eurofins Discovery’s state-of-the art Hit-Finding platform, our experts designed an optimized screening cascade combining cutting-edge High-throughput Screening (HTS) technologies with the latest Biophysics techniques. With advanced technologies, such as the Echo® MS and RapidFire MS for HTS, and SPR, MST, TSA, nanoDSF and ITC for Biophysics, the cascade is effective for primary screening, orthogonal assays and evaluating Mechanism of Action (MoA). There are many biophysical approaches which bring value to drug discovery programs, and it is the target properties which drive the selection of the most suitable technology. In this presentation, we will highlight the novel Spectral Shift technology, one of the techniques employed by the Dianthus (well-known for the MST-TRIC). The goal was to rapidly implement an assay that monitors the ability of PIM3 to bind to ATP, as well as to compounds and fragments in solution. Using the Echo MS’ acoustic-droplet ejection, nanoliter dispensing in 394-well plates, robust data by Spectral Shift were quickly generated for the fragment-based screening campaign. Hits were confirmed in Dose-response with µM-range affinity values (KD), and TSA was used as an orthogonal assay for the identification of stabilizers among the selected PIM3 binders. For a defined set of 826 proprietary fragments, a single dose screening at 600µM using Spectral Shift was run, leading to the identification of 11% of potential binders. The 100 fragments were then confirmed in dose-response titration and 80 binders were confirmed. The top 30 binders were selected for orthogonal characterization using TSA. This led to the selection of the top 6 confirmed fragments, on which a MoA study in MST-TRIC was performed. Following this evaluation, 3 allosteric binders were identified, with KD from 355 to 550µM (affinity of orthosteric binders from 55 to 333µM). Biophysics plays an important role in drug discovery by qualifying valuable assets and increasing the success rates of future Hit-finding programs. The fragment hits selected in this PIM3 Kinase project, which show ADP-Glo™ activities, are now ready for crystallography studies (NMR &amp; X-ray co-crystallization). Citation Format: Celine Legros, Vanessa Porkolab, Maud Sigoillot, Olivier Mirguet. Biophysics in kinase drug discovery programs: Spectral shift technology from fragment screening to mechanism of action [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 3094.

Abstract 6091: Multi-cancer early cancer detection and cancer-type identification with high sensitivity non-invasive using urine: Pancreatic lung cancer early detection

Abstract N-NOSE, a breakthrough cancer detection test from HIROTSU BIO SCIENCE, utilizes the unique chemotactic behavior of the nematode C. elegans. This nematode is drawn to the urine of individuals with 15 different types of cancers (including stomach, colon-rectum, lung, breast, pancreas, liver, prostate, uterus, esophagus, gallbladder, bile duct, kidney, urinary bladder, ovary, oropharynx) while avoiding urine from healthy persons, making N-NOSE a comprehensive primary screening tool. It has a high accuracy rate, with 87.5% sensitivity and 90.2% specificity for early-stage cancers. Being non-invasive and reasonably priced, N-NOSE has been a significant success in Japan since its 2020 launch, with over 500,000 screenings.This presentation delves into the technology aspect of N-NOSE in Japan, comparing its clinical research and key findings with competitors to underscore its effectiveness in early cancer detection. Furthermore, N-NOSE now advances to not only detect cancer but also identify its type in a noninvasive way, using urine and with high PPV.With pancreatic cancer being particularly lethal, we outline “N-NOSE plus pancreas,” the world’s first pancreatic cancer detection test at an early stage. This involves genetically modifying C. elegans to react specifically to early-stage pancreatic cancer urine samples, a development centered on the newly identified cr-4 GPCR in AWC neurons. Next, we profile our newest liver-cancer-specific test, “N-NOSE plus liver,” which offers early liver cancer detection with high sensitivity and specificity using urine.Finally, we highlight how N-NOSE bridges the gap in cancer screening and type identification, offering a complete, non-invasive solution. Citation Format: Eric Di Luccio, Takaaki Hirotsu. Multi-cancer early cancer detection and cancer-type identification with high sensitivity non-invasive using urine: Pancreatic lung cancer early detection [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 6091.

Abstract 1275: In vivo functional CRISPR screens identifies metabolic dependencies mediating triple negative breast cancer lung metastasis

Abstract Metastasis is responsible for about 90% of cancer-related mortality. Throughout the metastatic cascade, cancer cells requires substantial metabolic rewiring to adapt the nutrient micro-environmental changes enabling dissemination and seeding in distant organs. Recent insights into importance of the tumor-intrinsic metabolic reprograming has been implicated in immune escape, yet altered immune responses imposed by tumor metabolism during metastasis are poorly understood. Here, we generated genetic perturbations in triple-negative breast cancer cells using metabolism-focused CRISPR knockout library and performed in vivo screens in both immunocompetent and immunodeficient mice. By comparing enriched sgRNAs in lung metastases or primary tumors, we identify loss of one gene related to long chain fatty acid (LCFA) β-oxidation (FAO) potentiates metastatic dissemination in immunocompetent mice but no in immunodeficient mice, suggesting that impaired FAO in tumor cells promotes breast cancer lung metastasis through an lymphocytes-dependent manner. We confirmed individual sgRNAs targeting this FAO gene from the primary screen dramatically accelerate metastasis in different breast cancer orthotopic models. Specifically, we find that low expression of this gene in primary tumors of advanced breast cancer patients with high grade tumors and positive lymph nodes (LN+) is associated with reduced overall survival. In addition, deletion of this gene generates an immunosuppressive microenvironment in the lung metastatic tumors with increased neutrophils, monocytes and decreased TNFα+-IFNγ+ CD8+ T cells. Mechanistically, impaired FAO in tumor cells causes mitochondria imbalance under lipid metabolism stress, consequently triggers inflammatory response on establishing a neutrophil-rich metastatic niche via mitochondrial nucleic acid-driven innate immunity. Altogether, our data provides a new insight on how tumor cell lipid metabolism impacting immune interplay during metastatic progression, nominating potential anti-tumor immunotherapies. Citation Format: Xiaoyong Wang, Shu-ting Chou, Verra Ngwa, Yoonha Hwang, Breelyn Karno, Deanna Edwards, Jin Chen. In vivo functional CRISPR screens identifies metabolic dependencies mediating triple negative breast cancer lung metastasis [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 1275.

Abstract 86: Deep immuno-profiling of syngeneic tumor mouse models for preclinical studies

Abstract The identification of the major cellular players involved in the progression of a type of cancer is a key step for the success of new immunotherapies for personalized medicine. Immune cells play critical functions in cancer, and mice with intact immune systems are vital to understanding tumor immunology. It is however a daunting challenge as complex relationships interplay between tumor cells and the immune system. Each component of innate immunity or adaptive immunity, such as T lymphocytes, macrophages or neutrophils, may be directed to a pro- or anti-tumor function. In order to cope with the complexity of the tumor microenvironment, it is necessary to use an experimental approach capable of characterizing the heterogeneity of the cell types present in the tumor, the evolution of their relative proportion and their fine-tuned functional specificity. This approach leads to the identification of new cellular biomarkers of different stages of tumor progression in order to identify new targets for therapeutic time-window of and improve cancer diagnosis To decipher the impact of immunotherapy treatments involved in the anti-tumor response, cellular phenotyping of leukocytes infiltrating a tumor but also those present in peripheral organs is necessary. Essentially based on extracellular labeling, this primary screen aims to quantify the different cell populations present in a syngenic tumor models on B6/N or BalbC genetical backgrounds. In order to increase our understanding in the precise mode of action of anti-PD1 treatment at the cellular level in sensitive and unsensitive models, we investigated immunophenotypes and responses to immune checkpoint inhibitor (ICI) of several hallmark syngeneic tumor models (MC38, CT26, B16F10, B16-OVA, RENCA, EMT6) in immunocompetent mouse models by flow and mass cytometry. We compared growth kinetics and profiled the immune cell composition of tumor microenvironment (TME), draining lymph node (dLN) and blood in order to establish immune-phenotypic cell signatures that correlates with treatment efficacy. Supervised, unsupervised and integrative data analysis as well as visualization tools are used to identify significant changes across different experimental settings. Our results indicate that each model possesses a unique tumor-immune infiltrate profile that can be modulated with immunotherapies. Overall, these studies provide an important resource of highly-characterized syngeneic tumor model and highlight the importance of tumor immune landscape variance across models that will drive selecting the most appropriate model to test novel immunotherapeutic agents and enhance our translation of knowledge from syngeneic models to human tumors. Citation Format: Anais Joachim, Emilie Maturin, Marielle Mello, Lilia Hadjem, Magali Grange, Olivier Deas, Ana Zarubica, Bernard Malissen, Hervé Luche, François Romagne, Stéphanie Blanchin. Deep immuno-profiling of syngeneic tumor mouse models for preclinical studies [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 86.

Abstract 3097: CETSA HT: Employing rapid high-throughput target engagement technology in primary screening of P53

Abstract The confirmation of target engagement for candidate drugs is a fundamental requisite in drug discovery. A significant portion of projects fail to reach the clinic due to lack of efficacy or failure to show lead candidates interacting with the intended target in a more complex environment. The utility of using the CETSA® (CEllular Thermal Shift Assay) technology to measure target engagement in intact cells is increasingly exploited in high throughput screening, where examples demonstrate low false positive rates in screens of up to 500K compounds. The p53 tumor suppressor protein, a crucial guardian against cancer, functions as a transcription factor that mediates cancer suppression by regulating genes involved in a wide range of cellular processes such as DNA repair, cell cycle and apoptosis. Unfortunately, frequent mutations in the TP53 gene hinder p53's tumor suppressor role in human cancers, thus designating it as a promising target for anticancer therapy. However, attempts to restore p53 function for therapeutic purposes have not yet advanced to clinical approval, highlighting the complexities in leveraging p53 for cancer treatment. Its undruggable nature, attributed to the absence of accessible deep pockets, lack of enzymatic activity, and its intracellular location, further complicates targeted interventions with both low and high molecular weight drugs. Here, we demonstrate the analysis of a high-throughput plate based CETSA® HT screen on the challenging target p53, performed using AlphaLISA® SureFire® Ultra™ technology and our newly developed high-throughput screening platform. Hits identified from this screen are privileged from the onset by binding to the target in a physiologically relevant cellular matrix and in a binding-mode agnostic way. This is in contrast to traditional screening methods targeting activity or in vitro interaction where cellular activity is addressed in later stages. This assay cascade highlights a rapid target-to-hit lead generation strategy placing the emphasis on finding chemical matter that interacts with endogenous target in living cells first, followed by analysis of hit material activity. The ‘shortcut’ to chemical matter active in living cells represents significant potential cost savings in comparison to assay development time on recombinantly produced proteins and/or modified cell lines for traditional HTS screening approaches that still rely on target engagement assessment downstream. Citation Format: Merve Kacal, Tomas Friman, Laurence Arnold, Victoria Brehmer, Daniel Martinez Molina, Stina Lundgren. CETSA HT: Employing rapid high-throughput target engagement technology in primary screening of P53 [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 3097.

Abstract 1231: First-in-class AR-V7/AR-fl small molecule molecular glue degrader for prostate cancer treatment

Abstract Metastatic castration resistance prostate cancer (mCRPC) is a lethal disease due to the development of treatment to androgen receptor (AR) signaling inhibitors (ARSi) and taxane chemotherapy. Treatment resistance occurs partly due to the expression of AR splice variants that lack the ligand binding domain (LBD) and are constitutively active in the nucleus. AR-V7 is the most prevalent variant conferring clinical resistance to both ARSi and taxanes. Currently, there is no selective AR-V7 inhibitor leaving patients with limited therapeutic options. Thus, the development of selective AR-V7 inhibitors is a high priority, clinically unmet need. To identify AR-V7 pharmacologic inhibitors, we performed a high throughput small molecule phenotypic screen using enzymatic complementation and nuclear AR-V7 as the assay endpoint. Our primary screen of ~170K compounds (z score = 0.8), followed by a cell-toxicity counter screen and a secondary GFP screen identified hit compound 7907, as a dual AR-V7/AR-fl protein degrader, with unique chemotype compared to all known AR modulators. Hit to lead optimization by medicinal chemistry/SAR studies identified lead compound 15, with increased potency compared to 7907. Mechanistically, we showed that compound 15 shortened AR-V7 protein half-life by activating the ubiquitin-proteasome pathway and inducing proteasomal degradation of both AR-V7/AR-fl, without affecting their transcription. Importantly, compound 15 induced degradation occurred within 3hr of treatment and was blocked by the clinically approved proteasome inhibitor, bortezomib. TurboID proximity ligation assay identified distinct E3 ligases, uniquely interacting with AR-V7 or AR-fl. Using AR-V7/AR-fl deletion mutants we further showed that compound 15 activity is mediated by the N-terminal domain of AR, present in both proteins. Remarkably, compound 15 sensitized LNCaP95 cells (endogenous AR-V7/AR-fl expression) to enzalutamide suggesting potential therapeutic synergism and ability to reverse enzalutamide resistance. Ongoing studies aim to narrow down the binding site of compound 15 as well as identify the E3 ligases mediating its activity. Together, these data support a molecular glue degrader mechanism of action, consistent with published studies showing that molecular glue degraders are ideal for targeting classically “undruggable” proteins lacking an LBD or containing intrinsically disordered domains, as is the case for AR-V7. Currently, all AR-directed therapies target the LBD of AR-fl, inhibiting AR signaling. AR-V7 expression is a direct outcome of this inhibition, leading to AR-fl and AR-V7 co-expression in patient tumors. We posit that our drug candidate, by offering dual AR-V7/AR-fl inhibition in a single treatment, has the potential to not only benefit patients with mCRPC but also patients with hormone-sensitive disease, and delay AR-V7 expression. Citation Format: CheukMan C. Au, Catrina Estrella, Prerna Vatsa, Michelle Naidoo, Michael Miller, Peter T. Meinke, David M. Nanus, Paraskevi Giannakakou. First-in-class AR-V7/AR-fl small molecule molecular glue degrader for prostate cancer treatment [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 1231.

Abstract 3096: CETSA Navigate HT: A new paradigm for high-throughput screening

Abstract The CEllular Thermal Shift Assay (CETSA) technology is a powerful way to assess target engagement across various matrixes. By the introduction of the high-throughput format (CETSA Navigate HT), this technology offers a reliable high-throughput screening method in natural cellular environments that can successfully be applied in compound screening for hit identification and confirmation already during lead generation. Due to its broad applicability to native, full length protein targets in the live cellular environment, without the need for expensive and time-consuming protein and/or compound labelling efforts, CETSA Navigate HT may be regarded as a new paradigm for high-throughput screening and actives generation. As a case study to demonstrate the applicability of CETSA Navigate HT in hit identification, we have applied the technology for primary screening of a 12000 compound library for CDK4, a protein targeted in several cancers. Hits identified from this screen are privileged from the onset by binding to the target in a physiologically relevant cellular matrix and in a binding-mode agnostic way. This is in contrast to traditional screening methods targeting activity or in vitro interactions where cellular activity is addressed in later stages. Data from follow up studies using functional assays will be presented as well as mass spectrometry based CETSA selectivity profiling and comparison to commercially available de novo CDK4/6 inhibitors. Citation Format: Helena Almqvist, Victoria Brehmer, Laurence Arnold, Tomas Friman, Stina Lundgren, Daniel Martinez Molina. CETSA Navigate HT: A new paradigm for high-throughput screening [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 3096.

The association between organised colorectal cancer screening strategies and reduction of its related mortality: a systematic review and meta-analysis

BackgroundTo assess the long-term association between organised colorectal cancer (CRC) screening strategies and CRC-relate mortality.MethodsWe systematically reviewed studies on organised CRC screening through PubMed, Ovid Medline, Embase and Cochrane from the inception. We retrieved characteristics of organised CRC screening from included literature and matched mortality (over 50 years) of those areas from the International Agency for Research on Cancer in May 2023. The variations of mortality were reported via the age-standardised mortality ratio. A random-effects model was used to synthesis results.ResultsWe summarised 58 organised CRC screening programmes and recorded > 2.7 million CRC-related deaths from 22 countries where rollout screening programmes were performed. The CRC screening strategy with faecal tests (guaiac faecal occult blood test (gFOBT) or faecal immunochemical tests (FIT)) or colonoscopy as the primary screening offer was associated with a 41.8% reduction in mortality, which was higher than those offered gFOBT (4.4%), FIT (16.7%), gFOBT or FIT (16.2%), and faecal tests (gFOBT or FIT) or flexible sigmoidoscopy (16.7%) as primary screening test. The longer duration of screening was associated with a higher reduction in the pooled age-standardised mortality ratio. In particular, the pooled age-standardised mortality ratio became non-significant when the screening of FIT was implemented for less than 5 years.ConclusionsA CRC screening programme running for > 5 years was associated with a reduction of CRC-related mortality. Countries with a heavy burden of CRC should implement sustainable, organised screening providing a choice between faecal tests and colonoscopy as a preferred primary test.

Diagnostic Accuracy of the Primary Care PTSD for DSM-5 Screen (PC-PTSD-5) in Demographic and Diagnostic Subgroups of Veterans

BackgroundThe Primary Care PTSD Screen for DSM-5 (PC-PTSD-5) is a screening instrument designed to identify patients with probable PTSD and is mandated to be used in VA primary care settings. However, validation of the diagnostic accuracy of the instrument is lacking in various demographic and diagnostic groups.ObjectiveTo evaluate the diagnostic accuracy of the PC-PTSD-5 in demographically and diagnostically stratified groups of VA primary care patients.Design, Setting, and ParticipantsBased on a sample of 519 VA primary care patients (40.61% response rate), the PC-PTSD-5 was evaluated against a criterion measure, examining its sensitivity, specificity, and other diagnostic properties. Evaluations were conducted in demographically stratified patient groups, including gender, age, race/ethnicity, marital status, and education, as well as diagnostically stratified groups, in patients with and without a depression, alcohol use, and drug use disorder.Main MeasuresThe MINI-International Neuropsychiatric Interview was the criterion measure against which the PC-PTSD-5 was evaluated.Key ResultsAmong the 10 demographically stratified groups of patients, the PC-PTSD-5 demonstrated a sensitivity between 81.25% (95% C.I.: 54.35 – 05.95) and 100% (95% C.I.: 63.06 – 1000) and its specificity ranged from 80.54% (95% C.I.: 74.70 – 85.55) to 88.31% (95% C.I.: 83.45 – 92.15). Among the 6 diagnostically stratified groups of patients (with and without depression, alcohol use, and drug use disorders), the PC-PTSD-5 exhibited a sensitivity between 88.89% (95% C.I.: 65.29 – 98.62) and 95.92% (95% C.I.: 86.02 – 99.50), and its specificity varied from 60.00% (95% C.I.: 40.60 – 77.34) to 88.14% (95% C.I.: 84.50 – 91.19).ConclusionsThe PC-PTSD-5, suitable for a diverse range of VA primary care patients, shows satisfactory sensitivity and specificity across various demographic and diagnostic groups. Healthcare providers should be cautious of false positives in patients with depression or substance use disorders, given the potential symptom overlap with PTSD.

The yield of routine laboratory examination in osteoporosis evaluation in primary care

SummaryThis study evaluated the yield of routine laboratory examination in a large population of older women in primary care. The prevalence of laboratory abnormalities was low and the clinical consequences in follow-up were limited. There was a weak association of laboratory abnormalities with osteoporosis but no association with vertebral fractures and recent fractures.PurposeMost osteoporosis guidelines advice routine laboratory examination. We have investigated the yield of laboratory examinations in facture risk evaluation of elderly women in primary care.MethodsWe assessed the prevalence of laboratory abnormalities and their association with risk factors for fractures, recent fractures, low bone mineral density (BMD), and prevalent vertebral fracture in 8996 women ≥ 65 years of age participating in a primary care fracture risk screening study. In a sample of 2208 of these participants, we also evaluated the medical consequences in the medical records during a follow-up period of ≥ 1 year.ResultsVitamin D deficiency (< 30 nmol/L) was present in 13% and insufficiency (< 50 nmol/L) in 43% of the study sample. The prevalence of other laboratory abnormalities (ESR, calcium, creatinine, FT4) was 4.6% in women with risk factors for fractures, 6.1% in women with low BMD (T-score ≤ − 2.5), 6.0% after a prevalent vertebral fracture, 5.2% after a recent fracture and 2.6% in the absence of important risk factors for fractures. Laboratory abnormalities other than vitamin D were associated with low BMD (OR 1.4, 95%CI 1.1–1.8) but not with prevalent vertebral fractures nor recent fractures. Low BMD was associated with renal failure (OR 2.0, 95%CI 1.3–3.4), vitamin D insufficiency (OR 1.2, 95%CI 1.0–1.3) and deficiency (OR 1.3, 95%CI 1.1–.5). In the follow-up period, 82% of the laboratory abnormalities did not result in a new diagnosis or treatment reported in the medical records.ConclusionsWe identified a low prevalence of laboratory abnormalities in a primary care population of older women and the majority of these findings had no medical consequences.

Confirmatory Assay for Laboratory Diagnosis of Malaria Using Molecular Approach.

Prompt malarial treatment and surveillance is crucial for accurate diagnosis of Plasmodium Sp. Gold standard microscopic examination has been widely applied for diagnosis of malaria in most part of the endemic areas. But in case of submicroscopic and asymptomatic microscopic diagnosis is questioned. The study aims to develop a simple, cost effective & robust nucleic acid amplification technique for the detection of malaria parasite. Study population included 50 clinically diagnosed positive malaria patient samples from various pathological laboratories. Microscopy by preparing thick film was carried out of every sample for primary screening in the available facility of Surat Raktadan Kendra & Research Centre- Blood Bank. The conventional PCR (Polymerase Chain Reaction) was applied for genus-specific amplification targeting the 18S rRNA gene of Plasmodium. Agarose gel electrophoresis was used to separate and analyze the amplified PCR product using 2% Agarose gel. The study shows that nested PCR not only detected all microscopic positive samples, but also detected submicroscopic infections that were missed or misread by microscopy. Hence, the sensitivity of molecular based detection technique is proved to be more compared to microscopic examination.