Project Title: „ Interaction and dynamics of intestinal IgA and gut microbiome profile during antidiabetic therapy”
Funding: European Regional Development Fund (ERDF), Measure 1.1.1.1 “Support for applied research”
Project No.: 1.1.1.1/19/A/036
Period: 1 May 2020 – 30 April 2023
Project costs: 648 648 EUR
Principle Investigator: Dr. biol. Jānis Kloviņš
Cooperation partner: SIA Latvia MGI Tech
Project summary:
Type 2 diabetes (T2D) is a chronic disease affecting approximately 60 million people in the European Region that constitutes nearly 8.8% of the adult population. Despite its variable therapeutic effects, contraindications, and side effects, metformin is still the most frequently administered antidiabetic medication. It has recently become clear that metformin alters gut microbiota composition and may exert its function through these alterations, while mechanisms underlying this interaction are not fully understood. In our previous research, we have found that secretory immunoglobulin A (sIgA) may be involved in mediating this interaction. Therefore, we propose a hypothesis that sIgA is an important mediator of metformin effects on the gut microbiome and significantly modulates the T2D treatment.
The overall aim of the project is to understand the modulatory effect of sIgA on gut microbiome, specifically in case of T2D, and the role of metformin in this process by using clinical investigation in humans and corresponding T2D mouse model in order to identify reliable omics-based biomarkers for predicting the treatment outcome and to identify new treatment strategies.
We are planning to implement human studies to research this phenomenon using excellent clinical resource including ongoing OPTIMED trial and Genome Database of Latvian Population (LGDB), as well as mice studies. We will employ state of the art technologies including massive parallel sequencing based metagenome and transcriptome analysis as well as metabolite analysis of sorted microbiome components in conjunction with rich clinical information. The project will result in understanding not only the correlative associations between sIgA, microbiome, and T2D treatment efficacy but also functional mechanisms and causal interactions.
Information published 01.05.2020.
Progress of the project:
1 May 2020 – 31 July 2020
Involvement of patients with type 2 diabetes and healthy controls in the study and collection of necessary samples for further analysis of the intestinal microbiome are underway. The optimization of the bacterial cell sorting method after sIgA binding has been started.
Information published 31.07.2020.
Progress of the project:
1 August 2020 – 31 October 2020
The inclusion of patients with type 2 diabetes and healthy controls in the study and the collection of the necessary biological samples for further analysis of the intestinal microbiome are being continued. Optimization of bacterial cell sorting method after sIgA binding is performed, as well as preparation of biological samples for the functional analysis.
Information published 30.10.2020.
Progress of the project:
1 November 2020 – 31 January 2021
The involvement of patients with type 2 diabetes and healthy controls in the study and the collection of the necessary biological samples for further analysis of the intestinal microbiome and its fractions are being continued. The optimization of the bacterial cell sorting method after sIgA binding is being performed, as well as the preparation and processing of biological samples for functional analysis.
Information published 29.01.2021.
Progress of the project:
1 February 2021 – 30 April 2021
Involvement of the remaining cohort of type 2 diabetic patients and healthy controls in the study and collection of the necessary biological samples for further analysis of the intestinal microbiome and its fractions are ongoing. Bacterial cell sorting after sIgA binding is being performed, as well as preparation and processing of biological samples for analysis of microbiome composition and functional profile. Selection of mouse tissue samples for histological analysis has been started.
Information published 30.04.2021.
Progress of the project:
1 May 2021 – 31 July 2021
The involvement of the required T2D patients and healthy participants has been completed. The preparation and processing of the collected biological samples for the microbiome composition and its functional analysis continues. Selection of mouse tissue for histological analysis continues.
Information published 30.07.2021.
Progress of the project:
1 August 2021 – 31 October 2021
The preparation and processing of the collected biological samples for the microbiome composition and its functional analysis continues both in human and mice cohorts. Histological analysis of mice tissue continues. Work has begun on developing a mouse model and planning the course of the mice experiment.
Information published 29.10.2021.
Progress of the project:
1 November 2021 – 31 January 2022
The preparation and processing of the collected biological samples for the microbiome composition and its functional analysis continues both in human and mice cohorts. Data analysis workflows are being optimized; the obtained data is processed, visualized, and interpreted. Histological analysis of mice tissue continues. Work continues on creating a mouse model and planning the course of the experiment.
Information published 31.01.2022.
Progress of the project
1 February 2022 – 30 April 2022
The processing of samples obtained in the human cohorts, analyzes of shotgun-metagenome, metatranscriptome, and metabolome have been complete. Data processing, summarization, and interpretation have been performed. Histological analysis of mouse tissue is complete, data processing is ongoing. The analysis of metagenome and functional data in previously obtained mouse samples continues, as well as work on the development of the planned mouse model and planning of the course of the experiment.
Information published 29.04.2022.
Progress of the project:
1 May 2022 – 31 July 2022
The analysis of metagenome and functional data in previously obtained mouse samples continues, as well as work on the development of the planned mouse model and planning of the course of the experiment. The general taxonomic composition of the gut microbiome and the analyzed bacterial fraction coated with secretory IgA has been obtained.
Information published 29.07.2022.
Progress of the project:
1 August 2022 – 31 October 2022
Metagenome and transcriptome analysis in previously obtained mouse samples concluded. Correlation analysis of the obtained data in relation to the characteristics of the intestinal epithelium has been started. In parallel with the mouse model experiments, planning for functional studies of the interaction between IgA and metformin using in vitro bacterial cultures is underway.
Information published 31.10.2022.
Progress of the project:
1 November 2022 – 31 January 2023
Anaerobic bacteria cultivation from donor feces has been performed to assess the effects of metformin on bacterial growth in vitro, during experiments samples are collected for 16S rRNA sequencing (to assess changes in species composition induced by metformin) and for transcriptome sequencing (to asses metformin-induced changes in gene activity). In addition, a method is being developed to measure changes in the bacterial antigene affinity to the SIgA antibody caused by metformin. An experiment is underway in monocultures for taxons that were most abundantly represented in the SIgA-coated fraction from the results previously obtained in Type 2 Diabetes microbiomes.
Information published 31.01.2023.
Progress of the project
1 February 2023 – 30 April 2023
The cultivation experiment of anaerobic bacteria isolated from the donor’s faeces to evaluate the effect of metformin on bacterial growth, changes in species composition (16S rRNA sequencing) and gene activity (transcriptome sequencing) has been completed. Collection and processing of the obtained data is in progress. Development of a method for measuring changes in sIgA affinity caused by metformin has been carried out, measurements have been performed and data analysis is in progress. An experiment is underway in monocultures for the taxa that, from the previously obtained results, were most abundantly represented in the sIgA-bound fraction. Compilation and analysis of the first monoculture cultivation data was carried out.
Information published 02.05.2023.