Introduction

Many patients (pts) with ulcerative colitis (UC) do not achieve long-term remission with conventional therapies but are hesitant to switch to advanced therapies. Ozanimod (OZA) is an oral advanced therapy approved for the treatment of moderately to severely active UC. Durability of up to 3 years of continuous OZA treatment has been reported in the True North (TN) open-label extension (OLE).

Aims & Methods

The objective of the current analysis of the TN OLE was to assess the long-term durability of OZA treatment in pts who were inadequately controlled on conventional therapies at TN baseline. Advanced therapy–naive pts who were OZA clinical responders at TN Week 52 were grouped by baseline endoscopic disease activity as moderate (Mayo endoscopy subscore [MES]=2) or severe (MES=3). OZA efficacy (clinical remission, clinical response, corticosteroid (CS)-free remission, and endoscopic improvement) was evaluated at OLE W46 and OLE W94. Symptomatic clinical response (decrease from baseline in the combined 6-point rectal bleeding subscore [RBS] + stool frequency subscore [SFS] of ≥1 point and ≥30% and a decrease of ≥1 point in RBS or absolute RBS ≤1 point) and symptomatic clinical remission (RBS=0 and SFS ≤1 point [and decrease of ≥1 point from baseline SFS]) were assessed from OLE W5 to W94. All endpoints were assessed using observed case (OC) and nonresponder imputation (NRI) analyses.

Results

Of the 82 advanced therapy–naive pts who entered the OLE in clinical response after 52 wk of OZA treatment during TN, 45 had moderate baseline endoscopic disease and 37 had severe baseline endoscopic disease. Baseline demographic and clinical characteristics were generally similar across groups. Based on OC analysis, symptomatic clinical response and remission were observed in 100.0% and 90.5% of moderate pts, respectively, at OLE W5 and in 100.0% and 91.2% of severe pts, respectively, at OLE W5. Rates of symptomatic clinical response and remission were maintained through OLE W94. Rates of clinical remission, clinical response, CS-free remission, and endoscopic improvement at OLE W46 were similar in moderate and severe pts and were maintained through OLE W94 in the OC analysis (Table). Overall, the data followed similar patterns in the NRI analyses across endpoints.

Conclusion

Advanced therapy–naive pts who experienced disease progression on conventional therapies had sustained efficacy on OZA for up to 3 years. Baseline endoscopic disease activity did not affect the long-term durability of OZA responders. OZA may be an appropriate advanced therapeutic option for sustaining long-term symptomatic and clinical benefit in pts with UC after loss of response to conventional treatments.

Disclosure

This study was funded by Bristol Myers Squibb, Princeton, NJ, USA.
LPB: served as a speaker, consultant, and advisory board member for AbbVie, Amgen, Biogaran, Biogen, Boehringer Ingelheim, Celgene, Celltrion, Eli Lilly, Ferring, Forward Pharma, Genentech, H.A.C. Pharma, Hospira/Pfizer, InDex Pharmaceuticals, Janssen, Lycera, Merck, Mitsubishi, Norgine, Samsung Bioepis, Sandoz, Takeda, Theravance, Tillotts, and Vifor.
MDL: received research funding from Pfizer and Takeda; consulted for AbbVie, Bristol Myers Squibb, Calibr, Eli Lilly, Genentech, Janssen, Pfizer, Prometheus, Roche, Salix, Takeda, Target RWE, and Theravance.
BS: consulted for AbbVie, Arena, Bristol Myers Squibb, Boehringer Ingelheim, Celgene, Eli Lilly, Falk Pharma, Galapagos, Janssen, Pfizer, Prometheus, and Takeda; received speaker fees from AbbVie, CED Service GmbH, Falk Pharma, Ferring, Janssen, Novartis, Pfizer, and Takeda.
PI: received lecture fees from AbbVie, Bristol Myers Squibb, Celgene, Celltrion, Falk Pharma, Ferring, Galapagos, Gilead, Janssen, MSD, Pfizer, Sandoz, Sapphire Medical, Shire, Takeda, Tillotts, and Warner Chilcott; received financial support for research from Celltrion, MSD, Pfizer, and Takeda; received advisory fees from AbbVie, Arena, Boehringer Ingelheim, Bristol Myers Squibb, Celgene, Celltrion, Eli Lilly, Genentech, Gilead, Hospira, Janssen, MSD, Pfizer, Pharmacosmos, Prometheus Biosciences, Roche, Samsung Bioepis, Sandoz, Takeda, Topivert, VH2, Vifor, and Warner Chilcott.
FC: received research support from Takeda; consulted for Arena, Celgene, GlaxoSmithKline, and Takeda.
AJ, HW, and MTO: employees and/or shareholders of Bristol Myers Squibb.
DTR: received grant support from Takeda; consulted for AbbVie, AltruBio, Aslan, Athos, Bellatrix, Boehringer Ingelheim, Bristol Myers Squibb, Celgene, Chronicles, ClostraBio, Connect BioPharma, EcoR1, Eli Lilly, Genentech/Roche, Gilead Sciences, Iterative Health, Janssen, Kaleido Biosciences, Pfizer, Prometheus Biosciences, Reistone, Seres, Syneos, Takeda, Target RWE, and Trellus Health.
BA: received research funding from Takeda; consulted for AbbVie, Bristol Myers Squibb, Eli Lilly, Janssen, Medtronic, Pfizer, Samsung Bioepis, and Takeda; lectured for AbbVie, Bristol Myers Squibb, Eli Lilly, Janssen, Pfizer, and Takeda.

Introduction

The identification and establishment of reliable biomarkers and early diagnostic system for pancreatic ductal adenocarcinoma (PDAC) are urgently needed. Although blood-based liquid biopsy for comprehensive genomic profiling is useful for cancer diagnosis, we have recently shown that driver gene mutations derived from PDAC can be detected at a higher rate in pancreatic juice (PJ) than blood. We have also reported that some RNAs are carried within extracellular vesicles (EVs) and EV-encapsulated RNAs circulating in body fluid are useful as biomarkers for PDAC diagnosis. We recently developed a unique immunoprecipitation method called EViSTEP™ that utilizes a chelating-based reagent and EV surface marker antibodies (CD9/CD63) to isolate EVs with high purity and high yield.

Aims & Methods

In this study, we aimed to evaluate the usefulness of multi-layer liquid biopsy system by DNA mutation and RNA expression in PJ for cancer diagnosis. PJ samples (n=83; 15 benign hepatopancreatobiliary diseases, 33 low-grade (LG) IPMN, 10 high-grade (HG) IPMN and 25 PDAC) were collected during ERCP examination from each patient in a multicenter study. Cell-free (cf) DNA/RNA and EV-derived RNA (EV RNA) were purified from 2 ml of each PJ. Digital PCR was performed for driver gene mutation analysis (4 types of KRAS and 2 types of GNAS), as well as expression analysis of 10 long non-coding RNAs (lncRNA, e.g., HEVEPA, LINC02280) in cfRNA and EV RNA. The diagnostic performance of pancreatic cancer (PDAC+HG-IPMN) was statistically analyzed after normalizing RNA expression with two reference genes (ACTB/B2M).

Results

Logistic regression analysis (objective variables; PDAC+HG IPMN and others) with the KRAS and GNAS variant allele frequency (VAF), and RNA expression as factors was performed. The Area Under Curves (AUC) in Receiver Operating Characteristic (ROC) curve analysis for PDAC+HG IPMN diagnosis were 0.490-0.74 for KRAS mutations, 0.475-0.554 for GNAS mutations, 0.451-0.654 for cfRNA expressions and 0.362-0.639 for EV RNA expressions. Among them, the factors with relatively high AUC were KRASG12V (0.728), KRASG12R (0.687), cfRNA RP11-750H9.5/ACTB(0.654), EV RNA CRNDE/ACTB(0.633). The AUC values of the combinations, [KRASG12V, KRASG12R, cfRNA RP11-750H9.5/ACTB and EV RNA CRNDE/ACTB] and [the aforementioned four factors and CA19-9], were 0.876 and 0.885, respectively in terms of discriminating PDAC+HG IPMN. Moreover, concerning the diagnosis of Stage 0/I PDAC, the AUC values of EV HEVEPA, CA19-9, and the combination [EV HEVEPA and CA19-9] were 0.693, 0.746 and 0.84, respectively, suggesting that EV HEVEPA would be reliable biomarker for early detection of PDAC.

Conclusion

By incorporating cell-free lncRNA and EV lncRNA expression analysis into DNA mutation analysis in PJ samples could be expected to improve the diagnostic performance of pancreatic cancer. Liquid biopsy using PJ would predict the presence of neoplastic lesions with high probability and has a powerful potential for early diagnosis. Establishing the multilayer gene panel using these DNA and RNA has the potential to improve the early diagnostic accuracy for pancreatic cancer, a disease that exhibits significant diversity.

Disclosure

Grants for commissioned/joint research: Hitachi High-Tech Corporation (KT, YM), H.U. Group Research Institute G.K. (KT)

Introduction

A male patient in his twenties presented with clinical aspects of primary sclerosing cholangitis (PSC), chronic cholestatic liver disease, and hypothyroidism. PSC is a rare, chronic cholestatic liver disease characterized by biliary strictures and peribiliary fibrosis (1). Susceptible loci were identified in GWAS but the correlation between genetic background and disease pathogenesis needs further investigation (2). In the index patient we identified a heterozygote candidate variant in the ABCC6 gene (c.2279G>A, p.(Arg760Gln)). Dysfunctional ABCC6 was described to cause ABCB11 and ABCG5/G8 genes overexpression, leading to imbalanced bile composition resulting in gallstone formation and cholestasis (3). ABCC6 variants were previously described in cases with low phospholipid-associated cholelithiasis (LPAC) and intrahepatic cholestasis of pregnancy (ICP)(4-6) . Subsequently, we aimed to investigate rare variants in ABCC6 gene in whole exome sequencing (WES) results from patients with suspected cholestatic liver disease.

Aims & Methods

Variant evaluation and annotation analysis was carried out on WES data, with focus on loss of function and missense variants, presenting allele frequency (AF) of ≤1% based on gnomAD database (https://gnomad.broadinstitute.org). Variants were filtered to include only those in genes relevant to the patients’ clinical phenotype. For each rare variant, a comprehensive in silico assessment was performed using tools such as REVEL, AlphaMissense, EVE, SIFT, and MetaLR to evaluate potential pathogenicity. Filtered were prioritized and interpreted using ACMG-AMP guidelines. This systematic approach ensured a clinically meaningful and evidence-based evaluation of the identified variants.

Results

Nine different rare heterozygote variants were identified in fifteen samples of 247 WES results. Among these, the missense variants c.179G>A, p.(Arg60Gln) and c.3139C>T, p.(Arg1047Cys) were found in two LPAC patients. Additionally, the c.1171A>G, p.(Arg391Gly) variant was observed in two related cases: a mother with ICP and her child with elevated gamma-glutamyl transferase (GGT) levels and jaundice. Moreover, this variant was also detected in another patient with elevated transaminases. c.3871G>A, p.(Ala1291Thr) and c.955A>G, p.(Ile319Val) were seen in patients with ICP and cholestasis, respectively. Three other variants were also identified in patients with liver disease of unknown etiology (c.1540G>A, p.(Val514Ile) and c.2707G>C, p.(Glu903Gln) and c.4397G>A, p.CYS1466Tyr). Additionally one splicing variant (c.37-1G>A, p,?) was also detected in a patient with cholangiocarcinoma (CCA). All variants were observed in heterozygote state and classified as variants of uncertain significance (VUS).

Conclusion

An in-depth analysis of WES data led to the identification of several potentially disease-relevant variants in the ABCC6 gene. These variants may play a contributory role in the observed hepatic phenotypes, suggesting a broader functional involvement of ABCC6 beyond its well-established association with pseudoxanthoma elasticum (PXE). Our findings highlight the importance of integrating genetic data with detailed clinical phenotyping to enhance understanding of genotype–phenotype correlations and reveal novel roles for known genes in atypical or organ-specific presentations.

References

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4. Liu X, Lai H, Xin S, Li Z, Zeng X, Nie L, et al. Whole-exome sequencing identifies novel mutations in ABC transporter genes associated with intrahepatic cholestasis of pregnancy disease: a case-control study. BMC Pregnancy Childbirth. 2021;21(1):110.
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Disclosure

The authors declare that they have no conflict of interest.