Super-Obese Patient-Derived iPSC Hypothalamic Neurons Exhibit Obesogenic Signatures and Hormone Responses

Abstract

The hypothalamus contains neurons that integrate hunger and satiety endocrine signals from the periphery and are implicated in the pathophysiology of obesity. The limited availability of human hypothalamic neurons hampers our understanding of obesity disease mechanisms. To address this, we generated human induced pluripotent stem cells (hiPSCs) from multiple normal body mass index (BMI; BMI ≤ 25) subjects and super-obese (OBS) donors (BMI ≥ 50) with polygenic coding variants in obesity-associated genes. We developed a method to reliably differentiate hiPSCs into hypothalamic-like neurons (iHTNs) capable of secreting orexigenic and anorexigenic neuropeptides. Transcriptomic profiling revealed that, although iHTNs maintain a fetal identity, they respond appropriately to metabolic hormones ghrelin and leptin. Notably, OBS iHTNs retained disease signatures and phenotypes of high BMI, exhibiting dysregulated respiratory function, ghrelin-leptin signaling, axonal guidance, glutamate receptors, and endoplasmic reticulum (ER) stress pathways. Thus, human iHTNs provide a powerful platform to study obesity and gene-environment interactions.

Brain organoids: advances, applications and challenges

“…Defects in interneuron migration are observed in dorsal-ventral forebrain 'assembloids' generated from iPSCs derived from individuals with Timothy syndrome that harbor a mutation in the L-type calcium channel gene (Birey et al, 2017). Moreover, hypothalamic organoids containing nuclei-like clusters of neuropeptidergic neuron subtypes are promising models for studying metabolic disorders and obesity (Qian et al, 2016;Rajamani et al, 2018).…”

Cis-regulatory architecture of human ESC-derived hypothalamic neuron differentiation aids in variant-to-gene mapping of relevant complex traits

“…There is a lack of epigenomic data characterizing the genetic regulatory architecture of the developing and mature human hypothalamus, limiting our ability to translate studies into information directly relevant for disease 5 . Recently, improvements in embryonic and induced pluripotent stem cell differentiation strategies 1,6,7 have partially mitigated the need to study human hypothalamic neurons ex vivo.…”

“…There is a lack of epigenomic data characterizing the genetic regulatory architecture of the developing and mature human hypothalamus, limiting our ability to translate studies into information directly relevant for disease 5 . Recently, improvements in embryonic and induced pluripotent stem cell differentiation strategies 1,6,7 have partially mitigated the need to study human hypothalamic neurons ex vivo. As the precise regulation of hypothalamic development remains poorly understood, differentiating hypothalamic neurons from ESCs provides an opportunity to study these cells and their precursors over time, which should lead to greater understanding of the development of hypothalamic-governed traits and diseases.…”

Quantitative mass spectrometry for human melanocortin peptides in vitro and in vivo suggests prominent roles for β-MSH and desacetyl α-MSH in energy homeostasis

“…Post-mortem human brain samples provide a snapshot of peptide production, but cannot provide information on the dynamic regulation of POMC processing and secretion. To address these issues, we [31] , [32] and others [33] , [34] , [35] developed methods to generate hypothalamic neurons in culture from human pluripotent stem cells (hPSCs). Since these hypothalamic neurons can be produced at scale and are readily manipulated, they provide an unprecedented opportunity to study human POMC biology.…”