Understanding the potential of modulating Host-Microbiome-Glycan interactions (“the triangle of sweetness”) to tackle non-communicable diseases

Project Information

InteractHoMiG

Grant agreement ID: 101161110

DOI

10.3030/101161110

EC signature date 17 June 2024

Start date 1 September 2024

End date 31 August 2028

Funded under

• The European Innovation Council (EIC)

Total cost

€ 3 920 718,75

EU contribution

€ 3 920 718,75

Coordinated by

UNIVERSITEIT UTRECHT
Netherlands

Periodic Reporting for period 1 - InteractHoMiG (Understanding the potential of modulating Host-Microbiome-Glycan interactions (“the triangle of sweetness”) to tackle non-communicable diseases)

Reporting period: 2024-09-01 to 2025-08-31

Summary of the context and overall objectives of the project

Glycans, namely human milk oligosaccharides (HMOs), play crucial roles in the functions of the gut, metabolism, and immunity. While HMOs are vital in infant development and exhibit bioactive properties in adults, it is required to analyze and decode their functions in depth.
The current joint project by Utrecht University and Inbiose aims to identify novel glycosyltransferase enzymes, produce previously inaccessible HMOs, test glycan-microbe interactions in an ex vivo simulated gut microenvironment, decode direct and indirect effects of HMOs on the gut barrier in an in vitro cell model, and integrate the host-microbe-glycan interplay (“the triangle of sweetness”) in a human-microbial crosstalk (HuMiX) model. Thanks to powerful statistical methods, machine learning, and natural language processing techniques, we envision a thorough understanding about glycan structure-function relationship and predict their potential applications with stratifications based on gender, diet, and health conditions. We expect to validate at least one HMO alleviating impact of inflammation under two diets that is ready for a clinical trial. Cutting-edge Omics methods and data analysis pipelines will allow us to understand better the host-microbe-glycan triangle, and will provide both a wealth of research data and an expanded range of HMOs as research tools and potential therapeutics and/or preventives in human non-communicable diseases and food-related health conditions.
Our unique consortium capacities and a wealth of experience in glycoengineering, biosynthesis, bioinformatics, and noncommunicable chronic disease targeting will enable us to achieve the project goals. The project will create additional opportunities for Utrecht University and Inbiose in the further exploration and commercialisation of HMOs. We fully expect InteractHoMiG to be a milestone project for glycobiology research and prebiotic/probiotic/postbiotic/synbiotic R&D within the EU and worldwide.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

The InteractHoMiG project work plan is divided into 10 WPs designed to maximise the use of time and resources to achieve the proposed objectives and summarised in the PERT chart. Over the last year, several tasks have been successfully executed, resulting in significant progress and some achievements were obtained.
We expanded our glycosyltransferase library by more than 200 enzymes and produced the first set of glycans for initial gut barrier function screening using a newly optimized Caco-2 intestinal epithelial model. In addition, we developed a microbial strain capable of producing a complex mixture of HMOs mimicking human milk composition, advancing our disruptive innovation in the field of infant formula and dietary supplements.

A panel of 20 human milk oligosaccharides was screened using the optimized intestinal epithelial Caco-2 cell model. Barrier integrity was assessed by TEER (transepithelial electrical resistance) measurements, yielding promising results , particularly for several newly developed HMOs. In parallel, we conducted preparatory experiments to optimize microbial strains, exposure conditions and readouts for Caco-2 cells within the HuMiX 4.0 model, while also developing a high-throughput screening platform based on a Hollow-Fiber Membrane Gut-on-a-Chip model to investigate host-microbe-glycan interplay.

Preparatory work was carried out over the past year, including the setup of computational infrastructure arrangement, development of OMICS analysis pipelines, refinement of workflows, establishment of SIFR protocol and product requirements, and preliminary planning and brainstorming for in vivo experiments in anticipation of the Animal Ethical Committee approval. These activities guarantee us to rapidly initiate core tasks once the required products and data become available. In addition, we performed a systematic review to collect and evaluate existing knowledge on the ability of HMOs to modulate intestinal barrier integrity. The review focuses on sialylated, fucosylated, and acetylated HMOs, aiming to map their structure-function relationships in regulating intestinal barrier integrity.
For the Precision nutrition portfolio, the InteractHoMiG consortium contributed ideas to develop further and to implement the strategic plan by joining the steering committee and actively participating in working groups. Potential cross-project collaborations with other projects were identified and will be further explored in the near future. Details of the portfolio strategic plan are published in the Publications Office of the European Union website.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

During the last reporting period, InteractHoMiG obtained several scientific achievements, with potential scientific and economic/technological impact.
- Over 200 enzymes were screened on different acceptors for complex HMO synthesisrevealing diverse activity profiles and substrate specificities. These results could serve as a toolbox to assemble well-defined glycans, and constitute the first library of in-depth characterized glycosyltransferases.We synthesized 20 glycans to supportthe first in vitro screening using Caco-2 cells in WP4.
- The glycan library can be released as a research tool, and our novel HMOs can be transferred under agreement as products to key stakeholders (universities, research institutes, private industry research), interested in expanding their glycobiology research. As a first step, our HMOs are transferred to other projects within the EIC Precision Nutrition portfolio.Our team created novel concept strains capable of producing multiple HMOs simultaneously, covering most of the total oligosaccharide mass found in mother’s milk. This achievement paves the way for more optimal infant formulas that closely mimic human milk and opens the door for a completely new type of health ingredients. We consider this a breakthrough in Novel Food and precision fermentation technologies, creating access to a disruptive HMO product that is the closest in composition to human milk available in the industry.We optimized the intestinal epithelial Caco-2 cell model to combine mucus production and barrier integrity in a single system, promoting proliferation, differentiation, and functional maturation, thereby enhancing its physiological relevance. This optimized model enables detailed assessment of glycans on epithelial barrier integrity, mucus production, tight junctions, and cytokine responses, providing a robust platform for testing HMOs (in the current project), other dietary components or pharmacological interventions.

InteractHoMiG work packages