A combined microphysiological-computational omics approach in dietary protein evaluation

Abstract

Food security is under increased pressure due to the ever-growing world population. To tackle this, alternative protein sources need to be evaluated for nutritional value, which requires information on digesta peptide composition in comparison to established protein sources and coupling to biological parameters. Here, a combined experimental and computational approach is presented, which compared seventeen protein sources with cow’s whey protein concentrate (WPC) as the benchmark. In vitro digestion of proteins was followed by proteomics analysis and statistical model-based clustering. Information on digesta peptide composition resulted in 3 cluster groups, primarily driven by the peptide overlap with the benchmark protein WPC. Functional protein data was then incorporated in the computational model after evaluating the effects of eighteen protein digests on intestinal barrier integrity, viability, brush border enzyme activity, and immune parameters using a bioengineered intestine as microphysiological gut system. This resulted in 6 cluster groups. Biological clustering was driven by viability, brush border enzyme activity, and significant differences in immune parameters. Finally, a combination of proteomic and biological efficacy data resulted in 5 clusters groups, driven by a combination of digesta peptide composition and biological effects. The key finding of our holistic approach is that protein source (animal, plant or alternative derived) is not a driving force behind the delivery of bioactive peptides and their biological efficacy.

Bioengineered intestinal tubules as a tool to test intestinal biological efficacy of lettuce species

“…The construction, extracellular matrix (ECM) coating, seeding, and cultivation of the bioengineered intestinal tubule were described before and performed according to 36 . In short, a hollow fiber membrane (HFM) was guided in a custom-made 3-dimensional printed polylactic acid chamber.…”

Wild and domesticated lettuce species differ in intestinal biological efficacy

“…Image analysis was done in Fiji ImageJ version 2.0.3 as previously described (5). In short, a z-stack was transformed into a maximum intensity projection.…”

“…In contrast to most organ-on-a-chip models that are being applied in biomedicine and drug research, this bioengineered intestinal tubule was validated to assess the safety and e cacy of nutrition. During these assessments, the integrity of the intestinal epithelial barrier using membrane-associated complexes of zonula occludens-1 as a marker, overall cell viability, activity of the brush border enzyme alkaline phosphatase, which is essential for the nal steps of digestion, and the release of nitric oxide (NO) and cytokines as markers of the immune status were tested upon exposure to food proteins (5)(6)(7)(8).…”