(100c) Investigating the Role of Metabolism in Human Stem Cell Differentiation and Development Using Organoid Models

Organoids are in vitro models capable of recapitulating complex architectures of human tissues including skin, liver, pancreas, and brain. They are typically derived from human embryonic stem cells and induced pluripotent stem cells, and are renewable, perturbable, and scalable tools for investigating tissue development and disease. Control over the differentiation and patterning of organoids have traditionally relied on the specific timing and addition of subsets of morphogens and growth factors that mimic in vitro biochemical gradients. However, organoids still face several limitations. These include heterogeneity between and within organoids, incomplete derivation or maturation towards specific lineages or tissues, and aberrant cell types and cell states that suggest non-physiological cellular stresses. Prior results in the literature hint at metabolic conditions affecting organoid development. In particular, previous studies with embryoid bodies have shown that physiological glucose conditions can promote growth, while work in cerebral organoids has indicated typical culture conditions yield endoplasmic reticulum and glycolytic stress compared to in vivo tissues.

Our aim is to investigate the influence of glucose metabolism on germ-layer and lineage specification in human stem cell-derived organoids. Our studies with H1-derived cortical organoids showed that organoids grown in low glucose conditions form large Choroid Plexus (ChP) cysts within two weeks of culture whereas the organoids grown in high-glucose conditions are limited to cortical cell types. Organoids in low glucose conditions lacked the neural rosettes, that are intrinsic to cortical organoids and were positive for expression of ChP markers TTR, AQP1, and ZO-1 by immunofluorescence analysis. In addition, morphological differences in neuroepithelium development were observed in low vs high glucose conditions for H9-derived cortical organoids. These findings highlight the importance of glucose metabolism in lineage specification and suggest that nutrient concentrations are important knobs controlling organoid development.

This study will contribute to the understanding of the interdependence between metabolism and differentiation and help inform strategies for the improvement of in vitro human organoid models. It has the potential to uncover molecular components and mechanisms that convert metabolic changes to differentiation patterns. Furthermore, this can lead to advanced models for developmental diseases and reveal new therapeutic avenues.