Direct Reprogramming of Glioblastoma Cells into Neurons Using Small Molecules

Abstract

Glioblastoma multiforme, a type of deadly brain cancer, originates most commonly from astrocytes found in the brain. Current multimodal treatments for glioblastoma minimally increase life expectancy, but significant advancements in prognosis have not been made in the past few decades. Here we investigate cellular reprogramming for inhibiting the aggressive proliferation of glioblastoma cells. Cellular reprogramming converts one differentiated cell type into another type based on the principles of regenerative medicine. In this study, we used cellular reprogramming to investigate whether small molecule mediated reprogramming could convert glioblastoma cells into neurons. We investigated a novel method for reprogramming U87MG human glioblastoma cells into terminally differentiated neurons using a small molecule cocktail consisting of forskolin, ISX9, CHIR99021 I-BET 151, and DAPT. Treating U87MG glioblastoma cells with this cocktail successfully reprogrammed the malignant cells into early neurons over 13 days. The reprogrammed cells displayed morphological and immunofluorescent characteristics associated with neuronal phenotypes. Genetic analysis revealed that the chemical cocktail upregulates the Ngn2, Ascl1, Brn2, and MAP2 genes, resulting in neuronal reprogramming. Furthermore, these cells displayed decreased viability and lacked the ability to form high numbers of tumor-like spheroids. Overall, this study validates the use of a novel small molecule cocktail for reprogramming glioblastoma into nonproliferating neurons.

Direct cell-fate conversion of somatic cells: Toward regenerative medicine and industries

“…Most chemicals used in the direct reprogramming studies have positive or negative effects for various signal transduction pathways, e.g., Wnt (CHIR99021), 123)-129) cAMP (Forskolin), 123),125),126),128)-130) TGFO (RepSox, A83-01, SB431542), 123)-128),131) Ca 2D (ISX9), 126),129) Shh (Purmorphamine), 131), 132) Retinoic acid (TTNPB), 123), 127) JAK/STAT (AZ960, SP600125), 125),133) Notch (DAPT), 129) AMPK (dorsomorphin), 130) PCK (GO6983), 125) HIF (KC7F2), 133) MAPK (PD0325901, ZM336372), 124),133) O1-integrin (Pyrintegrin), 133) and Rho/ROCK (Thiazovivin, Y-27632) 125),131) signaling pathways, and epigenetic modifications, e.g., histone lysine deacetylation (Valproic acid), 42),123)-125),127), 131) histone lysine acetylation recognition (I-BET151), 126), 129) histone lysine methylation (Bix01294, Dznep), 124),127) DNA methylation (RG108), 124) and DNA demethylation (Vitamin C). 124) Although several types of cells, such as neuronal cells, hepatocytes, cardiomyocytes, and skeletal muscle progenitor cells, have been induced with chemicals, half of the reported studies used gene transduction in conjunction with the chemical treatments.…”

From Dermal Patch to Implants—Applications of Biocomposites in Living Tissues

“…In addition, cells also displayed the formation of spheroids within the scaffolds, indication the feasibility of using these constructs as drug screening models. The scaffolds were treated with a GBM-reprogramming cocktail (cocktail that has shown to reprogram GBM cells to neurons [69]). While elongated neurites were not seen, cells had impaired ability to proliferate throughout the scaffold and were not able to form cell-like spheroids.…”

Overcoming the Blood-Brain Barrier: Functionalised Chitosan Nanocarriers

“…Therefore, research on the potential therapeutic benefit of small agents is gradually emerging, especially in the context of modulation of the cellular redox state, involved in the initiation of several neurodegenerative disorders, as extensively discussed by Schiavone and Trabace, 2018 [ 6 ]. Moreover, a cocktail of small molecules, including forskolin, has also been investigated for its ability to reprogram glioblastoma cells into neurons [ 7 ]. Wang et al, [ 8 ] developed an in silico method to predict the permeability of small molecules towards the BBB using unbiased atomic molecular dynamics simulations.…”