Klotho inhibits neuronal senescence in human brain organoids

Abstract

Aging is a major risk factor for many neurodegenerative diseases. Klotho (KL) is a glycosylated transmembrane protein that is expressed in the choroid plexus and neurons of the brain. KL exerts potent anti-aging effects on multiple cell types in the body but its role in human brain cells remains largely unclear. Here we show that human cortical neurons, derived from human pluripotent stem cells in 2D cultures or in cortical organoids, develop the typical hallmarks of senescent cells when maintained in vitro for prolonged periods of time, and that moderate upregulation or repression of endogenous KL expression in cortical organoids inhibits and accelerates senescence, respectively. We further demonstrate that KL expression alters the expression of senescence-associated genes including, extracellular matrix genes, and proteoglycans, and can act in a paracrine fashion to inhibit neuronal senescence. In summary, our results establish an important role for KL in the regulation of human neuronal senescence and offer new mechanistic insight into its role in human brain aging.

Translation of Cellular Senescence to Novel Therapeutics: Insights From Alternative Tools and Models

“…Another 3D organoid model has been developed by cortical neurons that are differentiated from human iPSC were shown to represent typical features of senescent cells, such as increased SA-β-gal activity, p16/p21 expressions, and inflammatory cytokines ( Shaker et al, 2021 ). This senescent phenotype was also accompanied by the significant downregulation of Klotho, a type I transmembrane protein with anti-aging properties ( Kuro-o., 2011 ; Massó et al, 2015 ; Shaker et al, 2021 ).…”

Neural Epidermal Growth Factor-Like Like Protein 2 Is Expressed in Human Oligodendroglial Cell Types

“…Since little is known about the 10 murine NSC hub gene expression in NSC of the early developing human CNS, we next investigated their expression in rNSCs and cNSCs derived from human-induced pluripotent stem cells (hiPSCs). To this end, human WTC iPSCs ( Kreitzer et al, 2013 ) were differentiated into NEct or NMP using our published protocols ( Shaker M. R. et al, 2020 ; Shaker et al, 2021a ) to generate populations of rNSCs and cNSCs, respectively ( Figure 2A, B ). As expected, human rNSCs and cNSCs both expressed the pan-NSC markers SOX1, SOX2, and NESTIN ( Figure 2A, B ), while only NMPs expressed BRA-T, confirming their caudal identity.…”

“…To address this knowledge gap, we investigated NELL2 expression in human brain organoids generated from human ESC and iPSCs because brain organoids mimic the cellular make-up of early developing human brain, including NSCs, neurons, oligodendroglia, and astrocytes ( Shou et al, 2020 ). The cortical brain organoids we generated from human NEct cells with our previously developed protocol ( Shaker et al, 2021a ) exhibited the layered architecture of the developing cortex with multiple neurogenic zones ( Figure 3A, B ). Immunostaining of 4-week-old brain organoid sections with PAX6 and CTIP2 antibodies confirmed the successful generation of ventricular zone and cortical plate (cortical layer V), respectively ( Figure 3C ).…”

Metadichol® induced expression of neuronal transcription factors with Human fibroblast Dermal cells

“…Klotho gene (KL) expression, increased 10-fold after treatment with 1 ng Metadichol®. Klotho, a type 1 transmembrane protein, has been shown to inhibit TP53 and regulate cellular senescence by repressing the p53/p21 pathway [32]. Thus, klotho prevents aging in primary human broblasts [33] and is neuroprotective.…”