TPS246 Background: Dordaviprone (ONC201) is an oral small molecule that antagonizes dopamine receptor D2/3 and agonizes the mitochondrial protease ClpP, leading to activation of the integrated stress response and induction of TNF-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis. In preclinical murine models, ONC201 reduces the burden of colorectal adenomas and modulates inflammatory cytokine signaling, supporting its development as a chemopreventive agent in populations at risk for colorectal cancer (CRC). Despite endoscopic surveillance and non-invasive screening, individuals with familial adenomatous polyposis (FAP) or a history of multiple adenomas remain at elevated risk for CRC, underscoring the need for safe, mechanism-driven preventive strategies. Methods: This first-in-human, multi-center, open-label Phase I prevention study evaluates the safety and tolerability of ONC201 in individuals with FAP or multiple colorectal adenomas of unknown genetic etiology. Eligible participants include adults (≥18 years) at high risk for recurrent colorectal adenomas, defined as either a diagnosis of FAP or findings of >5 small adenomas (<1 cm) or ≥3 adenomas with at least one ≥10 mm on colonoscopy within the past 5 years, excluding those with a history of Lynch syndrome (HNPCC). Participants with ≥2 adenomas ≥5 mm identified during standard-of-care colonoscopy will undergo baseline tissue (adenoma and normal mucosa) sampling, with at least one polyp intentionally retained and accessible by flexible sigmoidoscopy for post-treatment sampling. Subsequently, participants will be assigned to an ONC201 dose level (120 mg, 375 mg, 500 mg) and frequency (weekly versus every 3 weeks) using a “Rolling Six” rule-based design, based on tolerability. Each dose cohort will be comprised of at least 5 participants, with a maximum of 6 participants, who will receive oral ONC201 for approximately 12 weeks. At the end of treatment, participants will undergo flexible sigmoidoscopy (or colonoscopy if clinically indicated), during which time the remaining polyp(s) and additional biopsies of normal colorectal tissue will be collected. The primary endpoint is the proportion of participants with unacceptable toxicity to ONC201. Secondary endpoints are to evaluate the mean changes in TRAIL expression in adenomas and normal mucosa. Exploratory endpoints include serum cytokine and immune profile changes and tissue-based markers of proliferation, apoptosis, stemness, and NK-cell infiltration. Enrollment is ongoing across U.S. academic centers, including Brown University, University of Michigan, Cleveland Clinic Foundation, Ohio State University, and Washington University in St. Louis. Funding is provided by NCI/Division of Cancer Prevention. Clinical trial information: NCT05630794 .

TPS10628 Background: The TAM01 trial of low dose (5 mg) tamoxifen (LDTAM) vs placebo was associated with significant improvement in risk for breast cancer in postmenopausal women with a hazard ratio of 0.30 along with a favorable side effect profile. Risk reduction in premenopausal women was less clear, with a non-significant hazard ratio of 0.73. Tamoxifen in premenopausal women can induce substantial increases in systemic estradiol and in preclinical studies upregulate endocrine resistance gene AGR2 . Both phenomena may impact LDTAM efficacy in premenopausal women. A pilot study (NCT00853996) of 20 mg/d of the SERM acolbifene in premenopausal women was associated with reduction in mammographic density, benign breast tissue Ki-67, and estrogen response gene expression (Fabian et al; Cancer Prev Res 2015) with no increase in AGR2 or vasomotor symptoms. Further studies of LDTAM and acolbifene in premenopausal women are warranted assessing change in imaging and benign breast tissue risk biomarkers with change in systemic hormones, ovarian reserve, and drug metabolites. Methods: NCT05941520 is a randomized, double-blind Phase II trial performed as part of the University of Michigan Early Phase Clinical Cancer Prevention (ClinCaP) Consortium part of the Cancer Prevention Clinical Trials Network (CP-CTNet) comparing 6 months of tamoxifen 5 mg and acolbifene 20 mg. Eligible participants are premenopausal women ≥35 without prior invasive breast cancer, but with ≥ 2-fold increased risk for the disease. The primary endpoint is difference in change in levels of AGR2 mRNA between the two arms. Secondary endpoints are within-arm change in an endocrine response gene index (ERGI), mammographic density, and MENQOL. Exploratory endpoints include within-arm change in benign breast Ki-67, ER, PR and AGR2 protein, association of baseline Anti-Mullerian Hormone (a measure of ovarian reserve) with 6-month serum estradiol and change in tissue estrogen responsive gene expression and AGR2 . Based on the preliminary data, mean log base2 (fold change, FC) of AGR2 in the acolbifene arm is assumed to be -1, which is tantamount to a 50% reduction. The estimated SD of the log2(FC) is 2.25. Assuming a log2(FC) of +0.6 in the low dose tamoxifen arm (50% increase), and the same SD as in the acolbifene arm, 36 evaluable subjects per arm are required to detect, with 80% power at an alpha = 0.03 (two-sided), a difference in FC of this magnitude between the two arms. Secondary endpoints are assessed via paired samples t-test or Wilcoxon signed-rank test. Target enrollment in this 4-site trial is 80 over 2.5 years. The protocol opened for accrual at the University of Kansas Medical Center in October 2024 and as of January 2025 is pending activation at the other sites. Clinical trial information: NCT05941520 .

PURPOSETranslation of diagnostic technology developed at the laboratory bench involves iterations of user feedback and design modifications. Learning about translational requirements early in the design process facilitates the development of feasible prototypes that have a better likelihood of implementation in global health settings. During our development of a portable system to detect human papillomavirus (HPV) for cervical cancer screening in India, we encountered and solved issues related to research translation. We report our findings to help others deploying diagnostic technology for global health.MATERIALS AND METHODSWe designed a point-of-care system to process patient samples and diagnose HPV infection in the cervix. We continually shipped components of the system from the United States to our collaborating team in India to assess component condition and usability at the local site. We simultaneously developed HPV isothermal amplification assays that were fit for purpose. Cervical brush samples were used in our portable system for functional validation.RESULTSWe found ideal transport methods to ensure component quality and reagent stability through the international shipment chain. Portable systems should be designed as simple as possible for correct usage at the local testing site. Usability tests drove our design improvements from 28% to 93% success rates. We demonstrated proof-of-concept functionality of our portable system for 13 cervical brush samples on-site, with a sensitivity of 100% and specificity of 88.9%.CONCLUSIONIssues related to technology transfer for global health settings manifest during distribution and deployment of prototypes. We identified several issues during our synergistic design process and report recommendations on the basis of our experience.

IntroductionUnderstanding the humoral immune response to HPV is important for understanding the natural history of infection and developing biomarkers for early detection of cervical cancer. This has been technically limited by HPV type diversity and challenges of high-throughput protein expression and display. This study aimed to profile the humoral immune response to the proteomes of 12 HPV types in women with or without abnormal cervical cytology undergoing cervical cancer screening.MethodsTo detect serum antibodies (Abs) against HPV, we developed custom HPV high-density diffusion-free nucleic acid programmable protein arrays (HD-NAPPA) displaying the proteomes of 2 low-risk (HPV6 and 11) and 10 high-risk (HR) HPV types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52 and 58). Arrays were probed with sera from women undergoing screening for cervical cancer, with normal (n=82) or abnormal (n=54) cervical cytology. HPV DNA testing and typing were done on cytology samples from all participants using an assay that detects 37 HPV types.ResultsAbs to any HPV protein were detected in 47.6% (95% C.I.: 36.5-58.8%) and 40.7% (95% C.I.: 27.9-54.9%) of women with normal and abnormal cytology, respectively and in 44.9% (95% C.I.: 36.4-53.6%) of all women. HPV16 DNA was the most frequently detected type (36.8%, 95% C.I.: 27.4-47.4%), however, Abs against HPV16 were remarkably the least frequently detected (7.4%, 95% C.I.: 3.8-13.5%). The most frequently detected Abs were against L1, in 30.1% (95% C.I.: 22.7-38.7%) of all women (31.7% and 27.8% of women with normal and abnormal Pap, respectively). Abs against E1 and E4 were the most (in 24.3%, 95% C.I.: 17.5-32.5%) and least (13.2%, 95% C.I.: 8.2-20.4%) frequently detected E-Abs in all women, respectively. Among all subjects with antibodies to either L1 or L2, 39.0% (95% C.I.: 24.6-55.5%) of those with L1 antibodies and 51.9% (95% C.I.: 32.4-70.8%) of those with L2 antibodies were positive for the antigen from only one HPV type.ConclusionOur findings shed light on the kinetics of HPV-specific humoral immunity in women with normal or abnormal cervical cytology and highlight the need for comprehensive immune profiling in different health and disease stages.

While ducal carcinoma in situ (DCIS) is considered to be pre-invasive, some patients will develop metastatic disease after a long disease-free interval. The prevailing dogma posits that invasive local recurrence is the source of subsequent metastasis, and thus the goal of DCIS therapy is the prevention of local recurrence. Recently, this paradigm has been called into question by the observation that some women develop metastatic disease in the absence of local recurrence or even following bilateral mastectomies, suggesting early cancer dissemination in some patients. If the presence of circulating tumor cells (CTCs) can be verified on some patients with pure DCIS, then dissemination may be occurring earlier than previously thought, suggesting that these patients might require additional monitoring or treatment. Here, we present a workflow to isolate and characterize CTCs from DCIS patients. Using a high throughput size based inertial focusing microfluidic device, the Labyrinth, we isolated and identified CTCs in 66.6 % (12/18) of DCIS patients with an average of 1.337 CTCs per five mL. Immunofluorescence staining and single cell qPCR of CTCs reveal mesenchymal characteristics of the cells that may contribute to their ability to migrate and metastasize. Preliminary targeted DNA sequencing revealed single nucleotide variations previously found in DCIS samples. Overall, this data supports the hypothesis that cancer dissemination is occurring in a subset of DCIS patients earlier than previously thought. Additionally, the molecular characterization of CTC in DCIS patients may provide important information on their biological characteristics and associated clinical behavior.

TPS4212 Background: To prevent invasive esophageal adenocarcinoma (EAC), endoscopic eradication therapy (EET) is used to remove its precursor, Barrett’s esophagus (BE) with dysplasia. EET combines endoscopic removal of visible lesions with radiofrequency ablation (RFA) of surrounding BE to achieve complete remission of intestinal metaplasia (CRIM) and complete eradication of dysplasia (CED) to halt progression to cancer. Unfortunately, EET has metaplasia recurrence rates of 12.4%/year; thus, adjunctive cancer interception agents are needed to maintain the gains of EET. Data from pre-clinical studies and patient tissues demonstrate that the Hedgehog (Hh) pathway regulates esophageal stem cell activity and cell fate determination (squamous versus intestinal). Post-RFA reactivation of Hh signaling is hypothesized to drive BE recurrence; however, current FDA-approved Hh inhibitors are expensive and toxic. The antifungal itraconazole inhibits Hh signaling and has demonstrated antitumor activity in multiple cancers. In addition, it inhibits VEGFR and PI3K-AKT pathways, which are critical to BE development and neoplastic progression. Given its safety and affordability, itraconazole represents a promising strategy to reduce BE recurrence and EAC risk. Methods: This randomized, phase 2b, double-blind, placebo-controlled trial will evaluate itraconazole’s efficacy in accelerating BE eradication. Participants with high-risk BE, defined as BE ≥2 cm with low/high-grade dysplasia or intramucosal/T1 adenocarcinoma, undergoing ablation will be enrolled. Participants will be randomized 1:1 to receive 300 mg of oral itraconazole or placebo for two weeks before and four weeks after their first 2 sessions of EET. The primary endpoint is time to CRIM, a surrogate for long-term BE recurrence, measured in days. Secondary endpoints include time to CED, 12-month BE recurrence rates, safety, tolerability, and correlations between itraconazole levels and patient-reported outcomes. We will enroll 74 patients (37 per arm). We shall use a two-sided log rank test for right-censored time to event analysis to assess differences. The sample size calculation is based on anticipated surviving proportions at specific study times. We assume that the surviving non-CRIM proportions of patients at 3, 6, 9, and 12 months in the control arm to be 0.8, 0.5, 0.2, 0.1, respectively, and in the treatment arm to be 0.5, 0.2, 0.1, 0.05, respectively. To detect this effect size with 80% power at 5% level of significance, we need 30 evaluable participants in each group (with a plan to enroll 37 per arm to account for attrition). A Cox model will adjust for demographic and clinical variables. If successful, this trial could establish itraconazole as a novel adjunct to EET, reducing BE recurrence and lowering EAC risk. Clinical trial information: NCT06732388 .

Colorectal cancer (CRC) is the second leading cause of cancer-related death in the United States. Currently, screening is the most effective method to reduce mortality from CRC. Chemoprevention, which aims to delay or even prevent the onset of CRC through eradication or prevention of precursor colonic adenomas, is an alternative strategy to address this problem. The imipridone Dordaviprone/ONC201 is a potential chemopreventive agent that has been shown to induce apoptosis in cancer cells through both upregulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and the integrated stress response. More recently, we have shown that ONC201 induces TRAIL and apoptosis in a dose-dependent manner in adenoma-derived organoids and reduces adenoma formation in the Apcmin/+ CRC mouse model. We hypothesized that ONC201’s analogues, ONC206 and ONC212, would similarly enhance human colonic adenoma cell death through both TRAIL-dependent and TRAIL-independent mechanisms. We therefore evaluated the effects of ONC206 and ONC212 in two adenoma-derived organoid lines: one developed from an adenoma from an individual with familial adenomatous polyposis (FAP) and the second from a sessile serrated adenoma (SSA). Sensitivity of the FAP and SSA organoid lines to ONC206 and ONC212 was assessed using CellTiter-Glo® luminescent cell viability assay. Organoids were plated in triplicates and treated with ONC206 and ONC212 (concentration ranging from 0 uM to 10 uM). Results were analyzed after 72 hours of incubation. Western blotting was performed on lysates generated from both FAP and SSA organoids treated with either DMSO, ONC206, or ONC212 for 24, 48, or 72 hours. Co-culture experiments were performed with the adenoma-derived organoids (dyed using blue CMAC dye) and human NK-92MI cells (dyed with green CMFDA dye) treated with either DMSO, ONC201, ONC206, or ONC212. Fluorescent images following administration of red live/dead dye were performed using DAPI, FITC, and cherry red channels on ImageXpress® at 0, 24, 48, and 72 hours. ONC206 and ONC212 demonstrated antineoplastic effects on adenoma-derived FAP and SSA organoids, with half maximal inhibitory concentrations (IC50) less than that of ONC201 in FAP and SSA organoids. As was noted in ONC201 studies, the mechanism of action of ONC206 and ONC212 appears to be mediated through modulation of multiple pathways including the TRAIL pathway (TRAIL and DR5 upregulation) and the integrated stress responses (ATF4 upregulation). Co-culture with NK cells revealed a significant increase in NK-mediated organoid cell death. The ability of ONC206 and ONC212 to enhance NK-mediated killing is being evaluated. Further analysis of the effects of ONC206 and ONC212 on stemness markers are currently underway and may reveal other potential biomarkers and chemopreventive strategies for ONC206 and ONC212. Alexis J. Lannigan, Jasper Chan, Lanlan Zhou, Varun V. Prabhu, Michael Dame, Dean Brenner, Jason Spence, Wafik S. El-Deiry, Alexander G. Raufi. TRAIL-inducing imipridones ONC201, ONC206, and ONC212 demonstrate anti-neoplastic effects in colonic adenoma-derived organoids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 2333.

Abstract Background: Colorectal cancer (CRC) is the second leading cause of cancer-related death inthe United States and screening is the most effective method to reduce mortality. Chemoprevention, which aims to prevent CRC through eradication or prevention of precursor colonic adenomas, represents an alternative strategy to address this problem. The imipridone dordaviprone (ONC201) has been shown to induce apoptosis in cancer cells through both upregulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and the integrated stress response. More recently, we have shown that ONC201 induces TRAIL and apoptosis in a dose-dependent manner and reduces adenoma formation in the Apcmin/+ CRC mouse model. We hypothesized that ONC201 would similarly enhance human colonic adenoma cell death through both TRAIL-dependent and TRAIL-independent mechanisms. We therefore evaluated the effects of ONC201 in two adenoma-derived organoid lines: one developed from an adenoma obtained from an individual with familial adenomatous polyposis (FAP) and the second from a sessile serrated adenoma (SSA). Methods: Sensitivity of the FAP and SSA organoid lines to ONC201 was assessed using CellTiter-Glo® luminescent cell viability assay. Organoids were plated in triplicates and treated with ONC201 (concentration ranging from 0 uM to 20 uM). Results were analyzed after 72 hours of incubation. Western blotting was performed on lysates generated from organoids treated with either 1.69 uM DMSO, 1.69 uM ONC201, or 3.38 uM ONC201 for FAP and 1.23 uM DMSO, 1.23 uM ONC201, or 2.46 uM ONC201 for SSA for 0, 24, or 48 hours. Co-culture experiments were performed with the adenoma-derived organoids (dyed using blue CMAC dye) and human NK-92MI cells (dyed with green CMFDA dye) treated with either 845 nM DMSO, 845 nM ONC201, or 1.69 uM ONC201 for FAP and 615 nM DMSO, 615 nM ONC201, or 1.23 uM ONC201 for SSA. Fluorescent images following administration of red live/dead dye were performed using DAPI, FITC, and cherry red channels on ImageXpress® at 0, 24, 48, and 72hours. Results: ONC201 demonstrated antineoplastic effects in adenoma-derived FAP and SSA organoids. Half maximal inhibitory concentrations (IC50) of ONC201 in FAP and SSA organoids were 1.69 uM and 1.23 uM, respectively. As was noted in rodent studies, the mechanism of action of ONC201 appears to be mediated through modulation of multiple pathways including the TRAIL pathway (TRAIL and DR5 were both upregulated) and the integrated stress response (ATF4 was upregulated). Co-culture with NK cells revealed an increase in NK-mediated organoid cell death. The ability of ONC201 to enhance NK-mediated killing is still being evaluated. Further analysis of the effects of ONC201 on stemness markers and on T-cell-mediated organoid cell killing are currently underway and may reveal other potential biomarkers and chemopreventive strategies for ONC201. Citation Format: Alexis J. Lannigan, Jasper Chan, Maximilian Schwermann, Lanlan Zhou, Varun V. Prabhu, Michael Dame, Dean Brenner, Wafik S. El-Deiry, Alexander G. Raufi. TRAIL-inducing imipridone ONC201/TIC10 demonstrates anti-neoplastic effects in colonic adenoma-derived organoids [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 4780.

Related Article from Trials and Tribulations of Interrogating Biomarkers to Define Efficacy of Cancer Risk Reductive Interventions

Supplementary Figure S2 from Phase IIa Clinical Trial of Curcumin for the Prevention of Colorectal Neoplasia

MAPRE1 protein expression in tissue microarray

PDF file - 96K, ELISA data for individual markers in newly diagnosed samples separated based on stage at the time of diagnosis compared to matched controls. Only significant differences are demarcated. * indicates p<0.05, ** indicates p<0.01.

PDF file - 112K, Workflow of experiments and sample cohorts described in the manuscript.

Supplemental Table S1. Clinical Characteristics of Samples

Case/control ratios of 22 proteins selected for antibody microarrays in pre-diagnostic CRC plasma samples from the Women's Health Initiative cohort

Normalized spectra counts of AK1, CLIC1, and SOD1 in eight CRC cell lines