Introduction

Crohn’s disease and ulcerative colitis, two forms of inflammatory bowel disease (IBD) require lifelong treatment and patient monitoring. Current predictors of relapse and therapeutic success have limitations, therefore there is a high unmet need to identify novel biomarkers in IBD. Cytokines may be utilized for this purpose in IBD therefore we aimed to assess and compare the inflammatory cytokine profile of the serum and mucosa of IBD patients and healthy controls in a Phase 2 clinical trial. Furthermore, the presence and predictive potential of the selected markers in fecal samples were assessed to identify Inflammatory bowel diseases (IBD) are defined by unregulated immune response leading to intestinal inflammation. Monitoring approaches for disease of the inflammation are mostly invasive, time-consuming and expensive. Therefore, a simple, rapid and non-invasive test, with ability to differentiate IBD from other gastrointestinal conditions and to monitor disease activity, will have important clinical implications. Our previous cytokine profiling identified that Plasminogen activator inhibitor type 1 (PAI-1) is expressed in IBD patients with active disease, but not in control.

Aims & Methods

Therefore, our aim was to assess the utility of PAI-1 in monitoring disease activity and therapeutic response of IBD patients in different biological samples. Serum, biopsy, and fecal samples were collected from 132 IBD patients and 30 controls without IBD. Inclusion criteria included change of therapy or initiation of new treatment for IBD patients. Organoid cultures (OC) were established from biopsies. Immunofluorescence (IF) staining was used to observe the presence and localization of cytokines. ELISA method was used to define the level of PAI-1 in the mucosa, serum and fecal samples. For the gene expression analysis qRT-PCR was used to observe the relative mRNA change of PAI-1 and other cytokines in the mucosa and OCs.

Results

The applied screening approach detected the presence of plasminogen activator inhibitor 1 (PAI-1) in active IBD. We demonstrated that the serum, mucosal and fecal PAI-1 concentration is elevated in IBD patients showing clinical and endoscopic activity. Additionally, in responder patients the initial PAI-1 level decreased significantly upon successful therapy, whereas it remained unchanged in non-responders. OCs derived from active IBD patients produced higher concentration of PAI-1 compared to the controls suggesting that epithelial cells could be a major source of PAI-1. We also showed that faecal PAI-1 selectively increases in active IBD patients, but not in other organic gastrointestinal diseases.

Conclusion

We successfully demonstrated in a Phase 2 clinical trial that serum, mucosal and fecal PAI-1 expression reflects to the disease activity in IBD, which could be used to disease monitoring and possibly therapeutic response. The correlation between the fecal PAI-1 level and the endoscopic activity promotes that it could be used as a novel non-invasive disease specific fecal biomarker in the IBD diagnosis.

Disclosure

LDN-051 is owned by Ladon Therapeutics Ltd.

Introduction

Precision medicine aims to expand upon the summary average treatment effects reported in clinical trials by providing more refined, individualised estimates of treatment effects. The phase IIb TUSCANY-2 study (NCT04090411), which examined the efficacy and safety of afimkibart (RO7790121/RG6631; previously PF-06480605/RVT-3101), an anti-TL1A antibody, in patients (pts) with moderate to severe ulcerative colitis (UC), previously reported clinically meaningful treatment effects at UEGW 2024.¹ Here, we report post hoc analyses of the relationship between baseline clinical characteristics and Week 14 efficacy in pts enrolled in TUSCANY-2.

Aims & Methods

Pts were randomised to receive subcutaneous afimkibart 50mg, 150mg, 450mg or matched placebo (PBO) monthly during induction. In this post hoc analysis, all afimkibart doses were pooled, and the endpoint of interest was clinical remission by modified Mayo score (mMS; defined as stool frequency subscore=0 or 1; rectal bleeding subscore=0; and endoscopic subscore [ES]=0 or 1) at Week 14. Our investigation of more patient-specific treatment effects was guided by the Predictive Approaches to Treatment effect Heterogeneity (PATH) statement.² We developed a regularised logistic regression model to predict the probability of Week 14 remission and to help examine heterogeneity of treatment effects based on the following baseline characteristics: mMS, C-reactive protein (CRP), serum albumin, Robart’s Histopathology Index (RHI), number of failed advanced therapy classes, corticosteroid use, disease extent, disease duration, age and sex. Baseline faecal calprotectin (FCP) was investigated separately due to the extent of missing data.

Results

The proportion of pts achieving clinical remission by mMS at Week 14 in the afimkibart pooled dose arm was 32.5% (65/200; 90% confidence interval [CI]: 27.3 to 38.1) compared with 13.3% (6/45; 90% CI: 7.1 to 23.8) of pts in the PBO arm (treatment delta: 19.2%; 90% CI: 7.6 to 27.9). Based on calibrated predictions from the multivariate model, all of the individual TUSCANY-2 pts (both treated with PBO and afimkibart) had anywhere between a 2–76% probability of Week 14 remission at baseline. The clinical characteristics included in the model explained ~10% of the variability in the observed outcomes. There was no evidence that the clinical characteristics were associated with heterogeneous treatment effects on the relative scale (odds ratios appeared to be constant; p-value >0.5). On the clinically meaningful absolute scale, the treatment delta in the top 50% of pts with a higher baseline probability of remission (27–76%) was 28.5% (90% CI: 8 to 42; 45.2% in afimkibart pooled dose arm vs 16.7%, in PBO arm), compared with a treatment delta of 7.6% (90% CI: –7 to 18; 18.8% in afimkibart pooled dose arm vs 11.1%, in PBO arm) in pts with a lower baseline probability of remission (2–27%). Lower baseline CRP, lower RHI and higher serum albumin contributed to a greater probability of remission and absolute benefit from treatment with afimkibart.

Conclusion

Pts with inflammatory bowel disease (IBD) have diverse baseline characteristics that are associated with variability of treatment outcomes, yet clinicians rely on reported averages from clinical trials to make individual treatment decisions. This analysis demonstrates how precision medicine approaches, that account for diverse characteristics, may improve interpretation of positive IBD clinical trial results. Integrating genetic and novel biomarkers alongside clinical features could help to better predict individual outcomes for pts with IBD.

References

1. Danese S, et al. Presented at UEGW 2024 (Oral presentation OP079).
2. Kent D, et al. The Predictive Approaches to Treatment effect Heterogeneity (PATH) Statement. Ann Intern Med 2020;172:35–45.

Disclosure

This study was sponsored by F. Hoffmann-La Roche Ltd. Third-party medical writing support, under the direction of the authors, was provided by Victoria Dagwell, MSc, of Ashfield MedComms, an Inizio company, and was funded by F. Hoffmann-La Roche Ltd.
LP-B reports consulting or advisory role/board from AbbVie, Abivax, Adacyte, Alimentiv, Amgen, Applied Molecular Transport, Arena, Banook, Biogen, Bristol Myers Squibb, Celltrion, Connect Biopharm, Cytoki Pharma, Enthera, Ferring, Fresenius Kabi, Galapagos, Genentech, Inc., Gilead, Gossamer Bio, GSK, IAC Image Analysis, Index Pharmaceuticals, Inotrem, Janssen, Lilly, Medac, Mopac, Morphic, MSD, Nordic Pharma, Novartis, Oncodesign Precision Medicine, ONO Pharma, OSE Immunotherapeutics, Pandion, Par' Immune, Pfizer, Prometheus, Protagonist, F. Hoffmann-La Roche Ltd, Samsung, Sandoz, Sanofi, Satisfay, Takeda, Telavant, Theravance, Thermo Fischer, Tigenix, Tillots, Viatris, Vectivbio, Ventyx and Ysopia; speakers’ bureau from AbbVie, Alfasigma, Amgen, Arena, Biogen, Celltrion, Ferring, Galapagos, Genentech, Inc., Gilead, Janssen, Kern Pharma, Lilly, Medac, MSD, Nordic Pharma, Pfizer, Sandoz, Takeda, Tillots and Viatris. JRA reports consulting or advisory role/board from Janssen, Pfizer, AbbVie, Vedanta, Genentech, Inc., Seres Therapeutics, Ferring, GSK, Merck, Bristol Myers Squibb, Roivant, Celltrion, TRXBio and Shattuck Labs; speakers’ bureau from AbbVie and Janssen. SD reports consultancy fees from AbbVie, Alimentiv, Allergan, Amgen, Applied Molecular Transport, AstraZeneca, Athos, Biogen, Boehringer Ingelheim, Bristol Myers Squibb, Celgene, Celltrion, Dr Falk Pharma, Lilly, Enthera, Ferring Pharmaceuticals, Gilead, Hospira, Inotrem, Janssen, Johnson & Johnson, Morphic, MSD, Mundipharma, Mylan, Pfizer, F. Hoffmann-La Roche Ltd, Sandoz, Sublimity, Takeda, Teladoc Health, TiGenix, UCB, Vial and Vifor; lecture fees from AbbVie, Amgen, Ferring Pharmaceuticals, Gilead, Janssen, Mylan, Pfizer and Takeda. PSD reports employment at Northwestern University; consulting or advisory role/board from AbbVie, Adiso, Alimentiv, Celltrion, Genentech/Roche, Geneoscopy, Janssen, Shattuck and Takeda; research funding from Takeda and Bristol Myers Squibb. AB reports employment and stockholder from Pfizer. DEC reports employment and stock ownership from Pfizer. EP reports employment, stock or other ownership interests and patents, royalties, other intellectual property from Pfizer. SN reports employment and stock or other ownership interests from Pfizer. MSV reports employment and shareholder from Pfizer. KH reports no conflict of interest. LU reports employment at Genentech, Inc. CS reports employment, stock or other ownership interests from Genentech/Roche. DB reports employment at F. Hoffmann-La Roche Ltd. KL reports employment and stock or other ownership interests from Genentech, Inc. BES reports honoraria for consulting from AbbVie, Adiso, Agomab, Alimentiv, Amgen, AnaptysBio, Arena Pharmaceuticals, Artugen, AstraZeneca, Biolojic Design, Biora, Boehringer Ingelheim, Boston Pharmaceuticals, Calibr, Celgene, Celltrion, ClostraBio, Equillium, Enthera, Enveda, Evommune, Ferring, Fresenius Kabi, Fzata, Galapagos, Genentech/Roche, Gilead Sciences, GSK, Gossamer Bio, Imhotex, Index Pharmaceuticals, Innovation Pharmaceuticals, Inotrem, Kaleido, Kallyope, Merck, Microbiotica, Mitsubishi Tanabe, Mobius Care, Morphic, MRM Health, Nexus, Nimbus Discovery, Odyssey, OSE Immunotherapeutics, Palisade Bio, Progenity, Prometheus Biosciences, Prometheus Laboratories, Protagonist, Q32 Bio, Rasayana, Recludix, Reistone, Sanofi, Sorriso, Spyre, Surrozen, Target RWE, Teva, TLL Pharmaceutical, Tr1X, Union and Ventyx; consulting/speaking fees from Abivax; consulting/speaking fees/other support from Lilly; research grants/consulting/speaking fees/other support from Bristol Myers Squibb, Lilly, Janssen, Pfizer and Takeda; research grants/consulting fees from Janssen; stock/stock options from Ventyx.

Introduction

Obefazimod is an oral small molecule that has demonstrated short- and long-term efficacy and good safety profile in patients with moderate-to-severe ulcerative colitis (UC) (1,2). Obefazimod modulates inflammation by upregulating a specific anti-inflammatory micro-RNA (miR-124). In humans, obefazimod is metabolized via UGT1A9 as a main active metabolite (N-Glu). We examined whether the pharmacodynamics of obefazimod with regard to miR-124 upregulation in blood of patients receiving obefazimod during a 16-week phase 2b induction study (EudraCT number: 2018-003558-26) were correlated with AUC and Cmax of the parent drug and its metabolite.

Aims & Methods

Absolute quantification (QuantaSoft Pro) of the miR-124 copy number was performed by using droplet digital PCR technology on 205 whole blood samples from 205 patients receiving obefazimod (25, 50, 100 mg once daily) for 16 weeks. For pharmacokinetics assessment (Cmax and AUC of obefazimod and its metabolite), blood samples were collected in all patients on week 16 of study treatment. The correlation between miR-124 upregulation and Cmax and AUC was tested with Spearman's non-parametric correlation analysis.

Results

Daily obefazimod treatment (25, 50, 100 mg) dose dependently increased the number of miR-124 copies in blood of UC patients (Table). miR-124 upregulation at week 16 in blood of obefazimod-treated patients was significantly correlated with AUC for obefazimod and its metabolite. The miR-124 upregulation showed a significant correlation with Cmax of N-Glu, while no significant correlation was observed with the parent compound. There was a dose-dependent correlation between miR-124 upregulation and Cmax of the metabolite.

Number of miR-124 copies - Correlation of miR-124 upregulation and PK parameters

Conclusion

The pharmacodynamics of obefazimod with regard to miR-124 upregulation in blood of patients receiving obefazimod for 16 weeks correlates significantly with AUC and Cmax of N-Glu, and to a lesser extent with the parent drug. These findings are indicative of a dominant effect of N-Glu on miR-124 expression, which is consistent with the longer half-life and higher exposure of the metabolite compared to obefazimod. The lack of significant PKPD correlation between miR-124 increase and PK values with the highest dose of obefazimod suggests a saturation effect.

References

1. Vermeire S, Sands B, Tilg H et al. ABX464 (obefazimod) for moderate-to-severe, active ulcerative Colitis: a phase 2b, double-blind, randomised, placebo-controlled induction trial and 48 week, open-label extension. Lancet Gastroenterol Hepatol 2022, 7 (11): 1024-1035
2. Vermeire S, Sands B, Biguenet S et al. ABX464 (obefazimod) in patients with moderate-to-severe ulcerative colitis: an Interim 48-week the safety and efficacy analysis from an ongoing 96-week open-label maintenance phase 2b study. UEG journal 2022, 10 (S8): p.340

Disclosure

Bruce E Sands reports consulting fees from Abbvie, Alimentiv , Amgen, Arena Pharmaceuticals, Artugen Therapeutics, Astra Zeneca, Boehringer Ingelheim, Boston Pharmaceuticals, Calibr, Celgene, Celltrion, ClostraBio, Equillium, Enthera, Evommune, Fresenius Kabi, Galapagos, Genentech (Roche), Gilead Sciences, GlaxoSmithKline, Gossamer Bio, Index Pharmaceuticals, Innovation Pharmaceuticals, Inotrem, Kaleido, Kallyope, Merck, Morphic Therapeutics, MRM Health, Progenity, Prometheus Biosciences, Prometheus Laboratories, Protagonist Therapeutics, Q32 Bio, Sun Pharma, Surrozen, Target RWE,Teva, TLL Pharmaceutical, Ventyx Biosciences; consulting and speaking fees from Abivax; consulting and speaking fees and other support from Lilly; research grants, consulting and speaking fees and other support from Bristol Myers Squibb, Janssen, Pfizer, Takeda; research grants and consulting fees from Theravance Biopharma; and stock options from Ventyx Biopharma.
Séverine Vermeire has received grants from AbbVie, J&J, Pfizer, Takeda and Galapagos; Séverine Vermeire has received consulting and/or speaking fees from: AbbVie, Abivax, AbolerIS Pharma, AgomAb, Alimentiv, Arena Pharmaceuticals, AstraZeneca, Avaxia, BMS, Boehringer Ingelheim, Celgene, CVasThera, Cytoki Pharma, Dr Falk Pharma, Ferring, Galapagos, Genentech-Roche, Gilead, GSK, Hospira, Imidomics, Janssen, J&J, Lilly, Materia Prima, MiroBio, Morphic, MrMHealth, Mundipharma, MSD, Pfizer, Prodigest, Progenity, Prometheus, Robarts Clinical Trials, Second Genome, Shire, Surrozen, Takeda, Theravance, Tillots Pharma AG, Zealand Pharma.

Introduction

Therapeutic goals in inflammatory bowel diseases (IBD) have undergone considerable change in recent years, with a shift of focus from simply achieving clinical remission. Patient-related outcomes (PROs) have also become increasingly important as adjunctive tools in assessing the efficacy of medical therapy from the patients’ standpoint. Health-related quality of life (HRQoL) questionnaires, correlate well with disease activity and can detect meaningful clinical changes in IBD patients who either suffer a relapse of disease or achieve remission [1,2].

Aims & Methods

We aimed to assess the utility of SIBDQ in the long-term follow-up of IBD patients compared to the conventionally used markers such as clinical indices, CRP levels and endoscopic activity scores. Patients were scheduled for evaluation every 12 months, but additional evaluations were performed as required in case of worsening clinical condition. The main data collected at each study visit included clinical aspects (disease activity, on-going medication, quality of life assessement-The Short Inflammatory Bowel Disease Questionnaire), endoscopic and histological findings.

Results

In a prospective setting, 176 patients were included in the study (65 Crohn’s and 111 UC patients) , with a mean follow-up time of 16 months and 1.3 study visits per patient. Clinical activity was associated with significantly lower SIBDQ (median 4.3 vs 5.7 in those without clinical activity, p<0.001 mann whitney u) and increased inflammatory burden ( median CRP levels 1,99 mg/L vs. 7.2mg/L, p<0.001 mann whitney u). SIBDQ scores were also significantly higher in patients showing mucosal healing compared to those with active ongoing inflammation (5.8 vs 4.9, p<0.001 MWU), while inflammatory burden was also significantly different between patients with and without ongoing mucosal activity (1.6 mg/L vs 4.2 mg/L, p< 0.001 MWU). SIBDQ significantly improved over time, with a median of 4.7, 5.4 and 5.8 at visits 1, 2 and 3 respectively (p<0.001 MWU), which was also consistent with a decrease in the overall inflammatory burden in the cohort (median CRP levels of 4.2, 3.4 and 2.5 mg/L respectively).

Conclusion

Clinical activity is associated with significantly lower quality of life and increased inflammatory burden. QoL scores are significantly higher in patients showing mucosal healing compared to those with active ongoing inflammation.

References

[1] Jowett SL, Seal CJ, Barton JR, Welfare MR. The short inflammatory bowel disease questionnaire is reliable and responsive to clinically important change in ulcerative colitis. Am J Gastroenterol. 2001 Oct;96(10):2921-8.
[2] Irvine EJ^​, Zhou Q, Thompson AK.The Short Inflammatory Bowel Disease Questionnaire: a quality of life instrument for community physicians managing inflammatory bowel disease. CCRPT Investigators. Canadian Crohn's Relapse Prevention Trial. Am J Gastroenterol. 1996 Aug;91(8):1571-8.

Introduction

Hand in Hand Colon was founded to mitigate, expedite, streamline and optimize a patient's journey after undergoing a colonoscopy that unearthed a potential carcinoma. The project bridges the gaps prompted by the involvement of interdisciplinary fields from colonoscopy to diagnosis, surgery, oncological treatment and ultimately, recovery. The project, spearheaded and managed by coordinating care nurses, guides patients through the thicket of uncertainty, both bureaucratic; "where do I go now?" and emotional as patients take on the role of "cancer patient".
Services require a maze of services which can lead to miscommunications and trouble understanding and navigating through the medical system. Patients will need to wait for their biopsy results, meet with a surgeon, oncologist, or both, undergo medical imaging as well as understand what is going on around them, medical terms why they are meeting with specialists, why are they getting tests done and where all this will lead them.
Each of these steps presents the patient with obstacles: Israel's imaging availability is one of the lowest in the OECD, backed up labs, doctors work with different HMOs and available and short-wait appointments are hard to come by.
The project offers one-one-one guidance to each patient following colonoscopy with a suspicious finding by a coordinating care nurse. The nurse will guide patients through medical necessities while also presenting availability and information, supporting and easing on the overwhelmed patient.

Aims & Methods

The project is a centralizing source for the patient, working as a one stop shop: expediting results and appointments, prioritizing and clarifying medical requirements and lowering anxiety levels.
Ultimately, our goal is to achieve diagnosis early as to move forward with treatment and increase the chance of a favorable prognosis while also providing patients with the option to ask questions, understand what's going on and feel, somewhat, at ease, therefore also increasing patient responsiveness.

Results

The project began taking on patients in March 2022. The following only relates to the project at Assuta Raanana. As of January 2023, 72 patients have been referred to the site's coordinating care nurse, of them 92% have undergone expedited imagery, 46% have already undergone surgery, chemotherapy or both, 100% of biopsy analyses were submitted within two weeks and 100% of patients were contacted and offered service by the appointed nurse.
Since the project began, coordinating care nurses have participated in group sessions with expert clinicians receiving training and support on dealing with distraught patients given life changing news. Soon, nurses will also receive group sessions to work through their own emotions regarding their exposure and day-to-day work with cancer patients to manage stress, burn-out, tension and sorrow.
The nursing team works under the belief that a gastroenterological nurse's job does not begin and end with their accumulated knowledge, nursing skills, technical abilities but that these can work to adhere to the 6 C's of nursing as put forth by the NHS: care, compassion, communication, competence, courage and commitment that all come together to provide patients with the best and highest level of care possible.

Conclusion

Following our experiences during the project's first year of operation, we believe that a patient's success and overall experience can be bettered through this project.

Introduction

Patient-reported outcome measures (PROMs) are recommended for assessing patient-centered outcomes and have been suggested as a tool for quality improvement (QA), which is crucial for IBD related outcomes and as clinical trials' endpoints. Real world use of IBD PROMS as a monitoring tool that enables quick access to caregivers, is still rare. Maccabi Healthcare Services (MHS), a state-mandated healthcare provider with over 2.6 million members, emphasizes patients' cooperation in improving their health and quality of life.

Aims & Methods

Aims: 1. To implement an IBD PROMS platform according to an operational process, which triggers a response from an IBD nurse and a multidisciplinary team, for routine HMO clinical use.
2. To compare the characteristics of responders and non-responders and study immediate actions.
Methods: We have used pre-existing research on the IBD PROMS platform(1) to implement validated questionnaires (IBD control PROMIS-10 measures and general health and nutritional items). Questionnaires translated into four languages were sent every 3 months to IBD registry members (defined by EPI-IRN criteria) via text messages. The calculated results and scores are directed to the electronic medical record (EMR) and are accessible to all caregivers. Digital alerts are sent to an IBD nurse in cases of "red flags" or low scores. A steering committee designed an actionable plan defining specific nurse responses for each red flag, and whether to consult with the treating physician, dieticians or social worker. A pilot study with patients from 6 gastroenterologists included 2 rounds of PROMS. Descriptive statistics were used to compare the responders to at least one questionnaire to non-responders for demographic and clinical characteristics, as well as for immediate reactions. An automated, language-specific, personalized messaging system according to patient response was accessible to the responders, with relevant information and general recommendations.

Results

Overall participation rate of the pilot study was 137/777(17.6%). Thirty one (23%) patients had red flags and 52(38%) had sub-threshold IBD-control scores. Opening rate of the personalized messaging system was 41%. Comparison between responders and non-responders is shown in Table 1. Responders tended to be significantly older and with a higher BMI. Medical treatment including biologics was not significantly different between responders and non-responders. More disease related laboratory tests were performed among responders after their initial response.

Table 1: Characteristics of the study population and comparison between responders and non-responders (Mean ±SD, unless stated otherwise)

Conclusion

IBD PROMS has been successfully assimilated and operated at MHS EMR by a multidisciplinary team, using a supporting personalized automated platform. Significantly higher rate of disease monitoring actions such as calprotectin tests was detected for responders. Responders did not necessarily have a more active or severe disease. The response rate should be increased via robust education and training.

References

Daniel Deutscher, Clara Weil, Gabriel Chodick, Sveta Tsukinovsky, Keith Bodger, Matti Waterman, Revital Kariv. Implementing electronic patient reported outcomes in inflammatory bowel disease: patient participation, score reliability and validity.
Health Qual. Life Outcomes. 2023 Jan 13;21(1):2 ^.

Disclosure

This project is supported by Takeda pharmaceutical by a quality improvement grant

Introduction

Ulcerative colitis (UC) is a chronic, non-specific inflammatory disease of the bowel of unknown etiology. This disease is characterised by high difficulty in cure, high recurrence and cancer rates, which seriously endangers the lives and health of our population. Taraxasterol (TA), a major constituent of the traditional Chinese medicine dandelion, has been reported in several studies to have good anti-inflammatory and antioxidant activities as well as anti-tumour effects.

Aims & Methods

However, the molecular targets and mechanisms of action of TA against UC have not been fully elucidated.The Swiss Target Prediction, PharmMapper databases were used to obtain relevant targets for the drug TA. Targets for UC were obtained from GeneCards, OMIM, PharmGkb, DrugBank databases. Targets of TA anti-UC identified by intersection of drug target and disease target. The relevant targets of TA anti-UC were uploaded to the String database and the resulting data were imported into Cytoscape 3.9.1 software to construct a protein-protein interaction (PPI) network of drug-disease targets. At the same time, the top 10 hub targets were screened using CytoHubba software. Gene ontology (GO) and Kyoto Encyclopedia of Genomes (KEGG) pathway enrichment analyses were performed on TA anti-UC targets using R software, and molecular docking was performed by Autodock Vina 1.1.2. The GEO dataset was used to analyze the differential mRNA expression of the TA anti-UC top 10 hub gene in UC patients and the normal population. Lipopolysaccharide (LPS)-induced intestinal epithelial cells (NCM460) were used to construct an in vitro cellular inflammation model and a Dextran Sulfate Sodium Salt (DSS) induced acute mouse colitis model. The pivotal targets and key signaling pathways were also validated by RT-qPCR and Western blotting.

Results

A total of 129 relevant targets of TA against UC were obtained, of which the top 10 hub targets were GRB2, PTPN11, SRC, PIK3R1, LCK, EGFR, MAPK1, IGF1, ESR1 and JAK2. The PI3K-Akt pathway was significantly associated with TA treatment of UC as identified by KEGG enrichment analysis. GEO dataset analysis demonstrated that EGFR and GRB2 were upregulated in UC patients, while SRC, PIK3R1 and JAK2 were down-regulated. In vitro experiments revealed that TA treatment reversed the expression of EGFR, GRB2, PIK3R1 and JAK2 differential genes. Furthermore, TA was able to up-regulate the mRNA expression of pro-inflammatory factor IL-10 and decrease the mRNA levels of anti-inflammatory factors IL-6, IL-1β and TNF-α in LPS-induced NCM460 cells. In vivo assay confirmed that TA significantly alleviated DSS-induced colonic inflammation in mice, including reversal of weight loss and shortening of colon length, reduction of DAI score in mice, and improvement of intestinal barrier disruption. In vivo and in vitro experiments have confirmed that TA also inhibits the activation of the PI3K/Akt pathway, thus exerting an anti-inflammatory effect.

Conclusion

TA may play a therapeutic role in UC by inhibiting the PI3K-Akt pathway. EGFR, GRB2, PIK3R1 and JAK2 may be the most relevant potential therapeutic targets for TA in the treatment of UC.