Institute Of Cancer Research Research Articles

Optogenetic Engineering of STING Signaling to Remotely Control anti-Tumor Immunity

The cGAS-STING signaling pathway is regarded as a promising target for cancer therapeutics development. Here we present an ultra-light-sensitive optogenetic device, designated LiSmore for light-inducible SMOC-like repeats, that remotely control STING activation and downstream gene expression in antigen-presenting cells to mount innate immunity. We demonstrate that noninvasive light-triggered STING modulation effectively promotes dendritic cell (DC) maturation and antigen presentation, allowing T cells to be effectively sensitized to engage tumor antigens, thereby photo-boosting anti-tumor immune response. Importantly, combining LiSmore with immune checkpoint blocker (ICB) synergistically enhances antitumor effcacy in an immunosuppressive lung cancer model, which otherwise remains largely unresponsive to ICB treatment. Our findings establish that targeted optogenetic activation of STING signaling allows for the remote control of DC functions by boosting anti-tumor immunity at the tumor sites. Hence, LiSmore sets the stage for the development of wireless optogenetic immunomodulatory therapies in a light-switchable manner. Acknowledgment: This work was supported by the Welch Foundation (BE-1913-20220331 to Y.Z.), the Cancer Prevention and Research Institutes of Texas (RP210070 to Y.Z.), and the National Institutes of Health (R01GM144986 to Y.Z., R01HL134780 to R01 Y.H., R01HL146852 to Y.H., and R01CA240258 to Y.H.). Images were created by Biorender.com. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Daily glucose pattern and physical activity levels in cancer survivors: preliminary results from remote monitoring

Background: Cancer survivors have elevated risk for type 2 diabetes. Increasing physical activity improves glucose control. However, this evidence is majorly based on blood glucose metrics (e.g., glucose or hemoglobin A1c) at a single point in time. Little is known about the effect of physical activity on blood glucose changes throughout a day in cancer survivors. Thus, we tested the hypothesis that higher physical activity levels is associated with a more favorable daily glucose pattern in cancer survivors in free-living settings. Methods: This is a preliminary analysis of an ongoing randomized controlled trial (Project KNOWN) to examine the effect of physical activity on daily glucose pattern via continuous remote monitoring. A total of 11 cancer survivors were included in this analysis (90% female, 90% white, average age 62.5 years [range 44-76 yr], 67% obese, and 75% hypertension). Participants wore a wrist-worn accelerometer (for daily steps) and a blinded continuous glucose monitor (CGM; for daily glucose averages [interstitial glucose was measured at every 15 minutes] and daily frequency of glucose spikes [defined as glucose level >140 mg/dL]) for two weeks. Multilevel regression model was used to account for multiple observations within each individual and to separate the within-person (WP) and between-person (BP) effects. The WP effect represents deviation from one’s own mean on any given day, and the BP effect represents the individual mean deviation from the study sample mean. To explore moderating effect on the relationship between physical activity and daily glucose, inflammatory biomarkers, blood lipids, blood pressure and body fatness (DEXA scan) were measured. Results: At the WP level, participants with elevated (>4 mg/L) systemic C-reactive protein (marker of inflammatory status), on the day when they had higher-than-usual step count, their glucose average was also lower than their usual level ( p=0.04). At the BP level, daily steps were negatively associated with daily frequency of glucose spikes in participants with less severe hypertension, lower % gynoid fat, and higher android/gynoid ratio than others in the study sample (all p<0.05). Further, daily steps were negatively associated with daily glucose average in participants with higher trunk-to-total fat ratio than others (p<0.05). Discussion: This preliminary study demonstrated the use of data from wearable sensors to analyze intra- and inter-person variability in daily glucose patterns in cancer survivors. Results suggested the impact of physical activity on daily glucose patterns might differ by a person’s physiological status. Future research can further characterize meaningful daily glucose dynamics from CGM data in cancer survivors to better understand how physical activity may have downstream effect on their health outcomes. American Institute for Cancer Research. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

A Comparative Study of Cardiac Toxicity Analysis in Hypo-Fractionated Vs Conventionalfractionated Adjuvant Radiotherapy in Post-Mastectomy Leftbreast Carcinoma

Background: Post-mastectomy adjuvant radiotherapy improves long-term survival by reducing the risk of loco-regional recurrence in breast cancer patients with high-risk features. Considering radiation therapy to the left sided chest wall heart is an important organ at risk. Hypo-fractionated radiotherapy is increasingly being used due to involvement of fewer treatment sessions and is almost radio-biologically equivalent to conventionally fractionated radiotherapy. Objective: This study was conducted to evaluate cardiac toxicities between hypo and conventionally fractionated radiotherapy. Methods: This quasi-experimental study was done at the National Institute of Cancer Research and Hospital, Mohakhali, Dhaka from January 2019 to December 2019, 60 patients were purposively assigned into two groups and each group included 30 patients. Arm A: received Conventional fractionated radiotherapy (50Gy/25 fractions over 5 weeks, at 2Gy/fraction). Arm B: received Hypo- fractionated radiotherapy (40.05 Gy in 15 fractions over 3 weeks at 2.67 Gy/fraction). Echocardiography (ECHO), Electrocardiography (ECG) was performed to find the cardiac toxicities. Results: The mean age of the patients in arm A was 50.03 ± 6.82 years and that of arm B was 48.08 ± 5.08 years. No significant statistical difference was found between the two groups regarding socio-demographic variables and clinical presentations. Most of the patients in both arms showed grade 0 cardiac toxicity (arm A: 70% and arm B: 80%) & few showed grade 1 cardiac toxicity (arm A: 20% and arm B: 13.3%) and three patients (10.0%) in arm A and two patients (6.66%) in arm B had shown grade 2 cardiac toxicities six months after treatment. However, this difference was statistically not significant (p >0.05). Before treatment, the ejection fraction (EF) of arm A and arm B were 63.3 ± 5.621 and 64.7± 4.991. Six months after treatment the EF of both arms decreased (arm A 60.2 ± 4.051 & arm B 60.9 ± 5.121) but the difference was statistically not significant (p >0.05). Conclusion: Hypo-fractionated radiotherapy in the adjuvant setting for the treatment of left breast cancer has no additional cardiac toxicity in comparison to conventionally fractionated radiotherapy while maintaining same efficacy. J Bangladesh Coll Phys Surg 2024; 42: 169-175

Treatment Outcome in Locally Advanced Rectal Carcinoma and Its Correlation with Hypo-Fractionated Radiotherapy and Concurrent Chemoradiotherapy

Introduction: Rectal cancer is the second most common type within the large intestine, posing significant health risks globally. In Bangladesh, it ranks high among diagnosed cancers. Improved diagnostic methods aid in early detection, which is crucial for effective management. Treatment typically involves a combination of surgery, radiotherapy, and chemotherapy. Preoperative chemoradiotherapy is a standard approach. However, newer methods like preoperative hypo-fractionated radiotherapy with surgery promise better outcomes, including reduced recurrence rates and enhanced survival, with potential cost and time savings. Aim of the Study: The study aims to evaluate and contrast the efficacy of hypofractionated radiotherapy and concurrent chemoradiotherapy in patients with locally advanced rectal carcinoma. Methods: The study, conducted at the Department of Clinical Oncology at Bangabandhu Sheikh Mujib Medical University Shahbagh, Dhaka, Bangladesh and the Department of Radiation Oncology at the National Institute of Cancer Research and Hospital (NICR&H) in Mohakhali, Dhaka, Bangladesh, aimed to compare treatments for locally advanced rectal cancer over 1.5 years (From January 2018 to June 2019). A total of 80 patients were divided into two groups; Group A received hypofractionated radiotherapy, while Group B received oral capecitabine alongside external beam radiotherapy. Inclusion criteria involved confirmed adenocarcinoma within specified stages, with ethical clearances obtained. Patients underwent evaluations before treatment and response assessments posttreatment. Treatment modalities were defined, focusing on radiotherapy and chemotherapy. Data were analyzed using SPSS software, employing various statistical tests. Results aimed to address study objectives while minimizing biases. Result: The study involved 80 patients, divided into Groups A and B, each with 40 individuals. Group A mainly comprised individuals aged 41-50 (45%), followed by 31-40 (25%). Group B had the highest proportion aged 31-40 (30%) and 41-50 (32.50%). Both groups showed male predominance (1.3:1). Moderately differentiated tumors were common, with Group A having more poorly differentiated tumors. Stage III was predominant pre-treatment (65% Group A, 70% Group B). Most patients presented with per rectal bleeding (80%) and alteration of bowel habits (56.25%). Response rates to treatment were similar between groups. Tumor downsizing was more frequent in Group B. Sphincter-sparing surgery was feasible in both groups with no significant difference. Conclusion: The study comparing preoperative hypofractionated radiotherapy (RT) to concurrent chemoradiotherapy (CCRT) for locally advanced rectal carcinoma found similar efficacy in tumor response, downsizing, and sphincter preservation. Statistical analysis showed no significant differences. Both approaches showed acceptable short-term outcomes, offering clinicians options based on patient and resource considerations.

Socio-demographic Determinants for Distant Metastases of Breast Cancer

Background: In Bangladesh, breast cancer continues to be the second-most common disease overall (12.5%) and the most prevalent cancer in women (27.4%). Patients with late-stage illness who present in low-income regions with distant metastases have a very poor prognosis. Objective: To determine the socio-demographic variables for breast cancer distant metastases in Bangladeshi women treated at a tertiary level hospital. Materials and Methods: This case control study was done from July 2017 to June, 2018 in National Institute of Cancer Research and Hospital (NICRH), Mohakhali, Dhaka. Results: A total number of 42 patients were taken as cases and another 84 patients without distant metastases were taken as controls. Mean age in case group was 42.67 ±9.27 years and in control group it was 45.13±9.60 years. The majority of the study participants in both the case (33.3%) and control (38.1%) groups had only completed kindergarten. In both the case group (71.5%) and the control group (85.7%), the majority of the women were found to be married. In both categories, the majority of the participants were sedentary workers. Additionally, Table 4 demonstrates that there is no statistically significant difference between the two groups with regard to co-morbidities, a positive family history, cigarette use, type of employment, and the existence of obesity (P > 0.05), stage of primary tumor (p=0.002, OR= 5.81; 95% CI: 1.85 – 18.15). Conclusion: Both the diagnosis and the course of treatment are delayed. Due to their lack of resources and lack of knowledge, many of them did not complete the course of treatment. Additionally, each of these factors contributes to the spread of breast cancer to distant areas.

L-Glutamine Substantially Improves 5-Fluorouracil-Induced Intestinal Mucositis by Modulating Gut Microbiota and Maintaining the Integrity of the Gut Barrier in Mice.

This study investigates the potential of glutamine to mitigate intestinal mucositis and dysbiosis caused by the chemotherapeutic agent 5-fluorouracil (5-FU). Over twelve days, Institute of Cancer Research (ICR) mice are given low (0.5 mgkg-1) or high (2 mgkg-1) doses of L-Glutamine daily, with 5-FU (50 mgkg-1) administered between days six and nine. Mice receiving only 5-FU exhibited weight loss, diarrhea, abnormal cell growth, and colonic inflammation, correlated with decreased mucin proteins, increased endotoxins, reduced fecal short-chain fatty acids, and altered gut microbiota. Glutamine supplementation counteracted these effects by inhibiting the Toll-like receptor 4/nuclear factor kappa B (TLR4/NF-κB) pathway, modulating nuclear factor erythroid 2-related factor 2/heme oxygenase 1 (Nrf2/HO-1) oxidative stress proteins, and increasing mammalian target of rapamycin (mTOR) levels, thereby enhancing microbial diversity and protecting intestinal mucosa. These findings underscore glutamine's potential in preventing 5-FU-induced mucositis by modulating gut microbiota and inflammation pathways.

Alan Hall. 19 May 1952—3 May 2015

Alan Hall was born in a mining town in Yorkshire in 1952. He studied chemistry at Oxford and then, after starting his PhD in biochemistry in Oxford, he completed it at Harvard in the USA, where he made important discoveries about the evolution of antibiotic resistance in bacteria. Following postdoctoral studies in the UK and Switzerland, he set up his independent research in London at the Institute of Cancer Research, where he began his work which opened up the field of study of the Ras-homologous family of small GTPase proteins. Alan made seminal discoveries in this field, which revolutionized cell biology in the UK and internationally. Alan's work made connections between cell migration, cancer and cell signalling and provided a mechanistic understanding of how many cell processes work at the molecular level. Alan helped to found the Laboratory for Molecular Cell Biology in London and became the director in 2001. In 2006 he moved to the Memorial Sloan Kettering Cancer Center in New York as chair of Cell Biology. He was awarded the Feldberg Foundation Prize in 1993, the Novartis Medal (UK Biochemical Society) and the Louis Jeantet Prize for Medicine in 2005, as well as the Canada Gairdner Prize in 2006. He was elected as a European Molecular Biology Organization member in 1994, as a Fellow of the Royal Society in 1999 and Fellow of the Academy of Medical Sciences in 2004.

Aqueous extract of Epimedium sagittatum (Sieb. et Zucc.) Maxim. induces liver injury in mice via pyroptosis

Ethnopharmacological relevanceEpimedium sagittatum (Sieb. et Zucc.) Maxim. has been used traditionally in Asia. It can dispel wind and cold, tonify the kidney, and strengthen bones and tendons. However, adverse effects of E. sagittatum have been reported, and the underlying mechanisms remain unclear. Aim of the studyThis study aimed to investigate liver injury caused by an aqueous extract of E. sagittatum in Institute of Cancer Research (ICR) mice and explore its potential mechanisms. Materials and methodsDried E. sagittatum leaves were decocted in water to prepare aqueous extracts for ultra-high performance liquid chromatography analysis. Mice were administered an aqueous extract of E. sagittatum equivalent to either 3 g raw E. sagittatum/kg or 10 g raw E. sagittatum/kg once daily via intragastric injection for three months. The liver weights and levels of the serum biochemical parameters including alanine transaminase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TBIL), and alkaline phosphatase were measured. Hematoxylin-eosin staining was performed for histopathology. Apoptosis was detected using the TUNEL apoptosis assay kit. IL-1β was detected using ELISA kits. Proteomics was used to identify the differentially expressed proteins. Western blot analysis was performed to determine the levels of proteins significantly affected by the aqueous extract of E. sagittatum. ResultsE. sagittatum treatment increased the liver weights and liver coefficients, and ALT and AST levels significantly increased (p < 0.05). A high dose of E. sagittatum significantly increased LDH and TBIL levels (p < 0.05). Ruptured cell membranes and multiple sites of inflammatory cell infiltration were also observed. No evidence of apoptosis was observed. IL-1β levels were significantly increased (p < 0.05). The expressions of PIK3R1, p-MAP2K4, p-Jun N-terminal kinase (JNK)/JNK, p-c-Jun, VDAC2, Bax, and CYC were upregulated, whereas that of Bcl-2 was inhibited by E. sagittatum. The expression of cleaved caspase-1 was significantly increased; however, its effects on GSDMD and GSDMD-N were significantly decreased. The expression levels of cleaved caspase-3 and its effector proteins GSDME and GSDME-N significantly increased. ConclusionsOur results suggest that the aqueous extract of E. sagittatum induces liver injury in ICR mice after three months of intragastric injection via inflammatory pyroptosis.

Methodological examination of the 2018 World Cancer Research Fund/American Institute for Cancer Research (WCRF/AICR) Score.

The objective of this study was to examine the impact of methodological changes to the 2018 World Cancer Research Fund/American Institute for Cancer Research (WCRF/AICR) Score on associations with risk for all-cause mortality, cancer mortality, and cancer risk jointly among older adults in the National Institutes of Health (NIH)-AARP Diet and Health Study. Weights were incorporated for each score component; a continuous point scale was developed in place of the score's fully discrete cut points; and cut-point values were changed for physical activity and red meat based on evidence-based recommendations. Exploratory aims also examined the impact of separating components with more than one subcomponent and whether all components were necessary to retain within this population utilizing a penalized scoring approach. Findings suggested weighting the original 2018 WCRF/AICR Score improved its predictive performance in association with all-cause mortality and provided more precise estimates in relation to cancer risk and mortality outcomes. The importance of healthy weight, physical activity, and plant-based foods in relation to cancer and overall mortality risk were highlighted in this population of older adults. Further studies are needed to better understand the consistency and generalizability of these findings across other populations.

Abstract LB111: CORSA study - Telomere length measurement of 5371 Austrian colorectal cancer and adenoma patients and healthy controls: Relationships to tumor stage, course of disease and to clinical, life style and health parameters

Abstract CORSA study - Telomere length measurement of 5371 Austrian colorectal cancer and adenoma patients and healthy controls: relationships to tumor stage, course of disease and to clinical, life style and health parametersPetr Hanak1, Michal Kroupa1,3,4, Uttarilli Anusha1, Ludmila Vodickova1,3,4 Andrea Gsur2, Stefanie Brezina2, Pavel Vodicka1,3,4 1 Department of Molecular Biology of Cancer, Institute of Experimental Medicine of the Czech Academy ofSciences, Videnska 1083, 142 20 Prague, Czech Republic.2 Center for Cancer Research, Medical University of Vienna, Vienna, Austria;3 Institute of Biology and Medical Genetics, First Faculty of Medicine, Charles University, Prague, Czech Republic 4Faculty of Medicine and Biomedical center in Pilsen, Pilsen, Czech Republic Presenting author: Petr Hanák (e-mail: petr.hanak@iem.cas.cz)Telomeres are important parts of human chromosomes, playing a pivotal role in maintenance of genomic integrity. If their function is perturbed, it may trigger many pathologies, including colorectal cancer (CRC). In the present study, comparison of assessed relative telomere length (RTL) and tumor stage, progression of the disease and various health status and lifestyle parameters is investigated. Relative telomere length was measured in samples of DNA from peripheral blood lymphocytes of CRC patients by monochromatic multiplex qPCR. Repetitive telomere motif TTAGG was amplified from specific primer pair simultaneously with the reference albumin gene, the second one amplified from another set of primers specific to this gene. The qPCR reaction is based on intercalating dye Syto 9, which does not distinguish specific target. A separation of amplification of one target from the other one is based on division in time of both amplifications and on different levels of annealing temperatures of primer pairs. Relative telomere length was assessed in all patient samples. Obtained data correspond to our previously observed values. Here, we observe significant decreasing in RTL in the order healthy controls, adenoma and CRC patients (controls vs adenoma patients P&amp;lt;0.0001, controls vs CRC patients P=0.0001). Detailed statistical analyses and assignment to all relevant groups including health and lifestyle parameters is currently in progress. Acknowledgement &amp; Funding: The study was funded by the Grant Agency of Czech Republic (21-27902S), National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union – Next Generation EU and Ministry of Health of the Czech Republic (NU22J-03-00028) Citation Format: Petr Hanák. CORSA study - Telomere length measurement of 5371 Austrian colorectal cancer and adenoma patients and healthy controls: Relationships to tumor stage, course of disease and to clinical, life style and health parameters [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 LB111.

Transcriptional, growth factors, components of the AKT/mTOR signaling pathway, receptors and ligands of programmed cell death expression in melanoma

Introduction. Melanoma is the most dangerous neoplasm of the skin, characterized by a malignant and aggressive course. Transcriptional and growth factors, components of the AKT/mTOR signaling pathway, receptors and ligands of programmed cell death are involved in significant processes of oncogenesis.Aim. To study the expression of components of the AKT/mTOR (mTOR – mammalian target of rapamycin) signaling pathway, transcription and growth factors, expression of AMPK, LC3B, programmed cell death 1 (PD-1), programmed death-ligand 1 PD-L1 and programmed death-ligand 2 (PD-L2) in skin and mucosal tumor tissues.Materials and methods. The study included 21 patients with a verified diagnosis of melanoma of the skin of various localizations and mucous membranes of the nasal cavity T1a–4bN0M0 (I–IV stages) and 18 patients with basal cell carcinoma of the skin of various localizations T1–4N0M0 (I–VIA stages), aged 45 to 72 years old, who were treated in the department of head and neck tumors of the Cancer Research Institute, Tomsk National Research Medical Center. The presence of tumor ulceration was determined by microscopy and registration of the true absence of the epidermis over the tumor or due to traumatization of the epidermis. Expression of components of the AKT/mTOR signaling pathway, transcription and growth factors, expression of AMPK, LC3B, PD-1, PD-L1 and PD-L2 in the tumor tissue was determined by real-time polymerase chain reaction.Results. An increase in the expression of 70 S6 kinase and VHL was found in melanoma tissues compared to basal cell carcinoma. At the same time, the presence of signs of ulceration was associated with a low level of c-RAF, nuclear factor kappa B (NF-kB) p50 and hypoxia-inducible factor 1 (HIF-1) matrix RNA (mRNA) against the background of an increase in the expression of the hypoxia-inducible factor 2 (HIF-2) transcription factor. The study of the molecular features of neoplasms in relation to the tumor thickness according to Breslow revealed the contribution of transcription and growth factors, the intensity of intracellular signaling processes, modification of the microenvironment, autophagy and neoangiogenesis.Conclusion. The molecular and biological features of melanomas associated with invasive tumor growth have been identified. An increase in the expression of 70 S6 kinase and VHL are characteristic of a malignant skin tumor. The presence of signs of ulceration and tumor invasion were associated with a change in the transcriptional characteristics of factors with the induction of key markers, oncogenesis, which contributes to the formation of the invasive potential of the tumor.

Deep learning-based cervical lesion segmentation in colposcopic images

Artificial intelligence assisted cancer detection has changed the ream of diagnosis precision. This study aims to propose a segmentation network using artificial intelligence for accurately segmenting the cervix region and acetowhite lesions in cervigram images, addressing the shortage of skilled colposcopists and streamlining the training process. A computational approach is employed to develop and train a deep learning model specifically tailored for cervix region and acetowhite lesion segmentation in cervigram images. A dataset acquired in collaboration with KIDWAI memorial cancer research institute is used for building the model. Cervigram images are collected for training and validation, and a deep learning architecture is constructed and trained using annotated datasets. The segmentation network based on efficientnet architecture and atrous spatial pyramid pooling is designed to accurately identify and delineate the target regions, with performance evaluation conducted using precision, accuracy, recall, dice score, and specificity metrics. The proposed segmentation network achieves a precision of 0.7387±0.1541, accuracy of 0.9291, recall of 0.7912±0.1439, dice score of 0.7431±0.1506, and specificity of 0.9589±0.0131, indicating its reliability and robustness in segmenting cervix regions and acetowhite lesions in cervigram images. This research demonstrates the feasibility and effectiveness of using artificial intelligence-based computational models for cervix region and acetowhite lesion segmentation in cervigram images. It provides a foundation for further investigations into classifying cervix malignancy using AI techniques, potentially enhancing early detection and treatment of cervical cancer while addressing the shortage of skilled professionals in the field

Risk Factors of Early Age Colorectal Cancer among Patients Attending at National Institute of Cancer Research &amp; Hospital

Background: Colorectal cancer (CRC) poses a significant public health burden worldwide, with a notable trend of increasing incidence among individuals aged 20 to 50 years. Understanding the risk factors associated with early-onset CRC is crucial for developing targeted prevention and intervention strategies, particularly in limited-resource settings. Objective: This study aimed to identify and analyze the risk factors contributing to the development of early-onset CRC in a population from a limited-resource country. Method: A comparative study was conducted among 178 CRC patients admitted to the National Institute of Cancer Research &amp; Hospital (NICRH), Dhaka. Patients were divided into two groups: Group-A (age 20-50 years) and Group-B (age &gt;50 years). Demographic and clinical data were collected, and statistical analyses were performed to determine associations between various risk factors and early-onset CRC. Results: The mean age of Group-A subjects was 43.6±7.1 years, significantly younger than Group-B subjects (59.2±9.3 years). Male patients constituted 69.1% of the cohort, with a statistically significant male-to-female ratio of 2.2:1. Higher BMI (≥25) was strongly associated with early-onset CRC (p&lt;0.01). Occupations such as business and middle socioeconomic status were significantly linked to early-onset CRC. Regular exercise (&gt;30 minutes daily) demonstrated a protective effect against CRC development. Consumption of red meat and fast food was associated with a higher risk of CRC, while regular consumption of vegetables was protective. Familial adenomatous polyposis (FAP) presence was significantly associated with early-onset CRC. Conclusions: Male sex, high BMI, specific occupations, dietary factors, and presence of FAP emerged as key risk factors for early-onset CRC. Encouraging regular exercise and promoting healthy dietary habits, particularly reducing red meat and fast-food intake, are crucial in mitigating the risk of early-onset CRC in similar populations.

Investigator choice of standard therapy versus sequential novel therapy arms in the treatment of relapsed follicular lymphoma (REFRACT): study protocol for a multi-centre, open-label, randomised, phase II platform trial

BackgroundRelapsed or refractory follicular lymphoma (rrFL) is an incurable disease associated with shorter remissions and survival after each line of standard therapy. Many promising novel, chemotherapy-free therapies are in development, but few are licensed as their role in current treatment pathways is poorly defined.MethodsThe REFRACT trial is an investigator-initiated, UK National Cancer Research Institute, open-label, multi-centre, randomised phase II platform trial aimed at accelerating clinical development of novel therapies by addressing evidence gaps. The first of the three sequential novel therapy arms is epcoritamab plus lenalidomide, to be compared with investigator choice standard therapy (ICT). Patients aged 18 years or older with biopsy proven relapsed or refractory CD20 positive, grade 1-3a follicular lymphoma and assessable disease by PET-CT are eligible. The primary outcome is complete metabolic response by PET-CT at 24 weeks using the Deauville 5-point scale and Lugano 2014 criteria. Secondary outcomes include overall metabolic response, progression-free survival, overall survival, duration of response, and quality of life assessed by EQ-5D-5 L and FACT-Lym. The trial employs an innovative Bayesian design with a target sample size of 284 patients: 95 in the ICT arm and 189 in the novel therapy arms.DiscussionWhilst there are many promising novel drugs in early clinical development for rrFL, understanding the relative efficacy and safety of these agents, and their place in modern treatment pathways, is limited by a lack of randomised trials and dearth of published outcomes for standard regimens to act as historic controls. Therefore, the aim of REFRACT is to provide an efficient platform to evaluate novel agents against standard therapies for rrFL. The adaptive Bayesian power prior methodology design will minimise patient numbers and accelerate trial delivery.Trial registrationClinicalTrials.gov: NCT05848765; 08-May-2023.EudraCT2022-000677-75; 10-Feb-2022.

Abstract 4416: Rapid, low-input, targeted NGS workflow for DNA methylation

Abstract DNA methylation is a fundamental epigenetic process that regulates human gene expression. In cancer, methylation changes promote proliferation networks and metastasis. Development of biomarkers for methylation will be enabled by flexible, fast, low-input next generation sequencing workflows. We describe the first Ion AmpliSeq™ Methylation targeted panel and protocol on the turn-key Genexus™ Integrated Sequencer as part an ongoing collaboration with Ontario Institute for Cancer Research (OICR) to detect and predict response in early stage breast cancer and improve diagnostics for Black and Asian women. A 2-pool Ion AmpliSeq™ Methylation Panel for Breast Cancer Research was developed as a demonstration of design, workflow, and reporting for targeted, low-input methylation assessment in a multiplex setting across a variety of samples sources including FFPE. The panel contains 327 amplicons and was designed to target both strands and 10ng DNA input into bisulfite conversion was used for controls. A complete workflow begins at bisulfite conversion and progresses through the Genexus™ Integrated Sequencer which combines library construction, template preparation, and sequencing into a single run. The bioinformatics pipeline provides DNA methylation calls on both Watson and Crick strands at single base resolution and methylated:unmethylated ratios for each CpG. The entire end-to-end workflow was completed in 2 days, including a full analysis software solution. The panel was evaluated using 2 control gDNA samples. The first had an expected average methylation state across all CpGs of &amp;gt;98% and the second &amp;lt;5%. An equal mixture of these two samples was also tested. The Methylation Panel for Cancer Research performed well on control samples. The single lane Loading, Total Bases, Final Reads, and Raw Read Accuracy were &amp;gt;92%, &amp;gt;1G, &amp;gt;14 Million, and &amp;gt;90% respectively. The workflow was also demonstrated on research FFPE cancer samples. This Methylation Panel protocol offers a 2-day, end-to-end workflow with high resolution, targeted and quantitative methylation analysis from DNA input as low as 10ng into bisulfite conversion. The option to design custom methylation panels for interrogation of targets of interest without the need for whole genome methylation is now available. &amp;lt;For Research Use Only. Not for use in diagnostic procedures. Early access materials were provided by Thermo Fisher Scientific™ to OICR in support of this abstract, but no other financial support. This study was conducted with the support of the Ontario Institute for Cancer Research through funding provided by the Government of Ontario. © 2023 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified.&amp;gt; Citation Format: Loni Pickle, Andrew Hatch, Gokhan Yavas, Melanie Spears, Louis Gasparini, Vida Talebian, Anna Ying-Wah Lee, Mathieu Lariviere, Jane Bayani, Seth Sadis, Jeffrey M. Smith. Rapid, low-input, targeted NGS workflow for DNA methylation [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 4416.

Abstract 229: Molecular-genomic characterization of metastatic gastric cancer organoids and establishment of organoid-immune cell co-culture model

Abstract Advanced gastric cancer (AGC) showed marked molecular heterogeneity with aggressive behavior and treatment resistance, especially AGC with ascites from peritoneal metastasis. Tumor organoids are valuable in vitro model for mimicking organ characteristics and are used for developing therapeutic strategies. Therefore, we aimed to understand the characteristics of organoids established from AGC patient-derived fluids. Also, we are in progress of setting up an organoid-immune cell co-culture model for drug screening of immune checkpoint inhibitors (ICI) for AGC. All established organoids from peritoneal and pleural fluid of AGC patients were authenticated through short tandem repeat (STR) profiling, comparing with their matched PBMCs. We compared the morphological characteristics using Operetta High Content Imaging (Perkin Elmer). Also, the in house targeted sequencing and RNA sequencing data were obtained from the genome database of the Song-Dang Institute for Cancer Research and it was verified using molecular methods. Then, we co-cultured organoids with healthy donor PBMCs that were pre-activated using anti-human CD3/CD28 Dynabeads (Gibco). 3D spheroids, rather than single cells, and the PBMCs were co-cultured in matrigel (Corning Inc.) for more than 3 days. The overall success rate of the organoid establishment was 53.8% (78/145), with different sources of peritoneal (54%, 61/113) and pleural fluids (53.1%, 17/32). The organoid success rate was various as in EBV positive (66.7%, 2/3) and dMMR (0%, 0/4). Based on the analysis of 91 patients eligible for histopathology, the success rate of HER2-negative (57.5%, 46/80) is higher than that of HER2-positive (36.4%, 4/11). The organoids were observed in a variety of morphologies categorized in 3 groups such as grape-like, compact and glandular structure. Based on genomic data of 31 organoids, we could subgrouping for TCGA subtypes and EMT subtype. RTKs were amplified in 10 organoids, including the amplification of ERBB2 (n=2), MET (n=4) and FGFR (n=4). And KRAS alterations including amplification (n=2) and G12A/D (n=2) mutation were also detected. Then, we established the condition for organoid-immune co-culture model with PBMCs with Dynabead activation for 3-10 days. We confirmed that higher dose of PBMCs killed the tumor cells with reduction of organoid size and partial disintegration. We established the proper PBMC dose and characterization to maintain the organoids co-culture for further drug (ICI) sensitivity testing. In conclusion, we evaluated phenotypic and molecular characteristics of gastric cancer organoids established from malignant peritoneal and pleural effusion. The established organoid-immune cell co-culture models can be utilized to evaluate a variety of drug screenings, including ICI and various combinations, helping in the selection of drugs for individual patient. Citation Format: Han Byeol Mun, juin Park, Jinsoo Jang, Jingmin Che, Tae Soo Kim, Woo Sun Kwon, Hyun Cheol Chung, Sun Young Rha. Molecular-genomic characterization of metastatic gastric cancer organoids and establishment of organoid-immune cell co-culture model [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 229.

Abstract 7142: Optimizing estradiol dimers through linker design: Enhancing anticancer efficacy by targeting microtubule dynamics

Abstract We investigated the development and anticancer efficacy of novel estradiol dimers (ED), targeting the dynamics of microtubules—an essential factor in effective cancer therapy. Employing copper-catalyzed azide-alkyne cycloaddition (CuAAC), two distinct series of ED variants were synthesized. The initial series included thirteen dimers, derived from 17α-ethinyl estradiol, incorporating varied five-atom linkers with central carbon, nitrogen, or oxygen atoms, and, in some instances, modified with benzyl groups containing hydroxy or methoxy substituents. These dimers were evaluated for their cytotoxic properties and impact on the cell cycle, revealing certain structures with substantial efficacy, compared with the original ED. The subsequent series, encompassing twelve variants, concentrated on structural alterations within the aromatic bridges linking the estradiol units. In this series, linkers of simpler composition and fewer substituents demonstrated increased selectivity and potency against cancer cell lines, particularly ED3 and ED5, which exhibited cytotoxicity levels comparable to established tubulin inhibitors. Both dimers series showed the ability to disrupt microtubule assembly and mitotic processes, confirmed through cell-based evaluations and in vitro tubulin assembly assays. Complementing these results, in silico modelling and binding free energy calculations exhibited a strong correlation with the demonstrated biological activities, especially in terms of binding affinity to the colchicine site. Collectively, these findings highlight the promise of estradiol dimers as potential anticancer agents and emphasize the critical role of linker structures in enhancing their biological effectiveness. Work was supported by the Ministry of Education, Youth and Sports of the Czech Republic through infrastructural projects (CZ-OPENSCREEN - LM2018130; EATRIS-CZ - LM2018133) and National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union - Next Generation EU. Citation Format: Petr Dzubak, Michal Jurášek, Jiří Řehulka, Soňa Gurská, Pavel Polishchuk, Pavel Dráber, Lukáš Huml, Pavel B. Drašar, Alexandra Ivanova, Olena Mokshyna, Lenka Hrubá, Marián Hajdúch. Optimizing estradiol dimers through linker design: Enhancing anticancer efficacy by targeting microtubule dynamics [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 7142.

Abstract 7191: Targeting IGF1R/INSR pathway with approved ALK inhibitors inhibits AKT signaling and overcomes proteasome inhibitor resistance in multiple myeloma

Abstract Background: Resistance to proteasome inhibitors (PI) bortezomib (BTZ) and carfilzomib (CFZ) is a major obstacle to the successful treatment of multiple myeloma (MM); thus, identification of novel therapeutic options is an unmet medical need. ALK inhibitors have been shown to exhibit anti-MM activity in monotherapy or in combination with CFZ. Our data showed that MM cells were negative for ALK; thus, we aimed to identify the mechanism of action of the ALK inhibitor ceritinib in MM cells and the mechanism of synergy between ceritinib and CFZ. Methods: A set of PI-naïve and PI-resistant MM cells was used in this study. Genome-wide CRISPR/Cas9-based screening using the Brunello library was performed in the AMO-1 cell line. Kinase inhibitor selectivity data were retrieved from ChEMBL, v30 database. RNA sequencing was used to determine expression changes after treatment, and RNA-seq data from patients included in the CoMMpass study were analyzed. Western blotting was performed to assess the levels of total and phosphorylated protein. Unbiased LC-MS/MS was performed to determine the effects of ceritinib and CFZ on the intracellular metabolites. An in vivo mouse model based on the orthotopic injection of AMO-BTZ cells into the femur of NSG mice was used to determine the effect of the drug combination in vivo. Results: Among approved ALK inhibitors, ceritinib has been identified as the most synergistic cytotoxic combination with CFZ in PI-naïve and PI-resistant MM cells. An in silico search of the ChEMBL database identified InsR and IGF1R as receptor tyrosine kinase that are inhibited by ceritinib. PI-naïve, PI-resistant cell lines, as well as MM patients were negative for ALK, but positive for InsR and IGF1R expression. CRISPR/Cas9-screening identified genes involved in the negative regulation of mTORC signaling (DDIT4, NPRL2/3, TSC1/2) and the transcription factor FOXO1 as the major resistance candidates to ceritinib. Subsequently, FOXO1 inhibition using a selective chemical inhibitor protected the cells from ceritinib-induced cytotoxicity. Next, ceritinib inhibited mTORC signaling and induced the expression of genes related to cell cycle arrest, which are downstream of FOXO1. Likewise, ceritinib, by targeting the InsR/IGF1R, impaired purine-pyrimidine and amino acid homeostasis, suggesting metabolic and proliferation shut-down and amino acid starvation due to downstream mTORC and Akt inhibition and FOXO1 induction. The combination of ceritinib and CFZ was superior to CFZ in PI-resistant MM in vivo and showed strong synergistic cytotoxicity in primary cells from MM patients progressing under or after PI-containing therapy. Conclusion: Ceritinib, an FDA-approved drug, overcomes PI resistance in MM by inhibiting InsR/IGF1R and downstream Akt signaling. Therefore, ceritinib is a promising treatment option for PI-resistant MM. Supported by the project National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union - Next Generation EU. Citation Format: Andrej Besse, Tiberiu Totu, Marianne Kraus, Anthonius P. Janssen, Jana Veprkova, Max Mendez Lopez, Ondrej Slaby, Marija Buljan, Mario van der Stelt, Christoph Driessen, Lenka Besse. Targeting IGF1R/INSR pathway with approved ALK inhibitors inhibits AKT signaling and overcomes proteasome inhibitor resistance in multiple myeloma [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 7191.

Abstract 817: Establishing a niche to improve Africa’s contribution to cancer research

Abstract Africa has been a key source of scientific discoveries to the global community, especially in areas of scientific experiments and implementation. However, Africans are underrepresented in their contribution to global science and within the African countries, access to adequate funding and infrastructure to do high quality science is limited. These inequities contribute to the 10/90 gap in which only an estimated 10% of global health research funding is used to address health problems in Africa. Thus, it is not surprising that today, Africa contributes just about 2 percent of world research output, 1.3 percent of research spending, and 0.1 percent of all patents. To bridge the aforementioned gaps, mitigate the effect of brain drain and structural constraints and improve support for the next generation scientists, the Nigerian Government recently established the National Institute for Cancer Research and Treatment (NICRAT). It is worthy to note that cancer incidence and mortality rates in Africa are increasing 1.1 million new cases, with 711,000 deaths reported in 2020. This indicate a mortality-to-incidence ratio (MIR) of 0.64. These cancer challenges in Africa necessitates a concerted approach towards effective cancer control and management. Here, we show in detail how the NICRAT support improving science while integrating with Nigerian national development plans and strategies. We also show how the NICRAT is establishing a mechanism for domestic funding, while fostering national and international collaborations for African scientists and encouraging women into careers in cancer research. Currently, the NICRAT’s key task also includes centralizing cancer control efforts, mobilizing funding from different sources for cancer research, and coordinating cancer treatment activities across Nigeria. In addition, the institute facilitates collaboration between researchers, healthcare providers, and patients, and provides guidance for cancer policy formulation and implementation. The establishment of NICRAT by Nigeria’s government as well as similar centers elsewhere in Africa may improve active participation of African scientists in scientific practice, discourse, and decision making and spotlight their intellectual contributions towards solving science-associated global problems. Citation Format: Usman A. Malami, Sani A. Malami, Uchechukwu E. Nwokwu, Usman W. Mohammed, Faruk Mohammed. Establishing a niche to improve Africa’s contribution to cancer research [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 817.

Abstract 3558: An integrated data platform to support the international alliance for cancer early detection

Abstract Cancer early detection is one of the most critical areas of cancer research, as it offers the greatest potential for improving patient outcomes. The International Alliance for Cancer Early Detection (ACED) is a global partnership of world-leading cancer research institutions in the UK and US, established in 2019 to accelerate and revolutionize research in this field. ACED brings together the expertise of the Canary Center at Stanford University, the University of Cambridge, the Knight Cancer Institute at Oregon Health and Sciences University, University College London, and the University of Manchester, together with Cancer Research UK, with the goal of catalyzing new collaborations and research in the field of early detection. One of the major challenges to enabling international collaborations across different institutions is the ability to collect, share and discover datasets between researchers and institutions. To enable collaboration on innovative data science, fundamental functions include, 1) controlled data sharing mechanisms; 2) structured metadata to enable data exploration and data discovery; and 3) enabling easy computational access to that data. To advance researchers' collaboration, ACED began development of an Integrated Data Platform (IDP). The ACED-IDP is based on the Gen3 software platform developed by the University of Chicago's Center for Translational Data Science. Based on software systems originally developed for the NCI's Genomic Data Commons, Gen3 has been used in several other data projects including The Blood Profiling Atlas in Cancer (BloodPAC), Australian BioCommons and numerous other research data platforms. The unique nature of ACED required unique innovations to be made for the development of the IDP. Each of the member institutions within the alliance has different existing computer infrastructures, separate authentication platforms and heterogeneous data types forming the basis of their research. The IDP sought a cloud-ready strategy, while still being cognizant of the extreme costs associated with cloud egress fees that hamper researchers' ability to download data. Additions were made to the Gen3 platform to allow for hybrid cloud support, allowing on-premises as well as cloud object storage systems to be linked to the platform. This innovation permits each institution to share files using the mechanisms that they see fit. To unify the various datasets, the standard Gen3 schema was replaced with one derived from the Fast Healthcare Interoperability Resources (FHIR) standard. To support import from clinical data sets, tooling to enable import and export Observational Medical Outcomes Partnership (OMOP) data has been integrated. With this platform in place, we hope to advance international collaborations and accelerate early cancer detection research. Citation Format: Brian Walsh, Liam Beckman, JD Burchett, Matthew Peterkort, Jordan Lee, Michael Fitzsimons, Peter Vassilatos, Binam Bajracharya, Craig Barnes, Jawad Qureshi, Robert Grossman, Carrie Yakura, Yaozhi Lu, Sarah Burge, Daniel Kelberman, Erin Watson, Kyle Ellrott. An integrated data platform to support the international alliance for 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 3558.