Investigation of 2-Hydroxypropyl-β-Cyclodextrin Treatment in a Neuronal-Like Cell Model of Niemann–Pick Type C Using Quantitative Proteomics

Abstract

Niemann−Pick, type C (NPC) is a fatal, neurovisceral lysosomal storage disorder with progressive neurodegeneration and no FDA-approved therapy. Significant efforts have been focused on the development of therapeutic options, and 2-hydroxypropyl-β-cyclodextrin (HP-b-CD) has emerged as a promising candidate. In cell culture, HP-b-CD ameliorates cholesterol storage in endo/lysosomes, a hallmark of the disorder. Furthermore, in animal studies, treatment with HP-b-CD delays neurodegeneration and extends lifespan. While HP-b-CD has been promising in vitro and in vivo, a clear understanding of the mechanism(s) of action is lacking. Utilizing a neuron-like cell culture model of SH-SY5Y differentiated cells and U18666A to induce the NPC phenotype, we report here a large-scale mass-spectrometry-based proteomic study to evaluate proteome changes upon treatment with these small molecules. In this study, we show that differentiated SH-SY5Y cells display morphological changes representative of neuronal-like cells along with increased levels of proliferation markers. Inhibition of the NPC cholesterol transporter 1 protein by U18666A resulted in increased levels of known NPC markers including SCARB2/LIMP2 and LAMP2. Finally, investigation of HP-b-CD treatment was performed where we observe that, although HP-b-CD reduces cholesterol storage, levels of NPC1 and NPC2 are not normalized to control levels. This finding further supports the need for a proteostasis strategy for NPC drug development. Moreover, proteins that were dysregulated in the U18666A model of NPC and normalized to control levels suggest that HP-b-CD promotes exocytosis in this neuron-like model. Utilizing state of the art mass spectrometry analysis, these data demonstrate newly reported changes with pharmacological perturbations related to NPC disease and provide insight into the mechanisms of HP-b-CD as a potential therapeutic.

Integrated multi-omics profiling highlights the benefits of resveratrol hydroxypropyl-β-cyclodextrin inclusion complex for A53T transgenic mice through the microbiota–gut–brain axis

“…16 As aforementioned, RHSD was more effective than an equal dose of RES alone, indicating that HP-β-CD as an excipient can enhance the biological activity and bioavailability of resveratrol. Interestingly, preclinical and clinical data suggest that HP-β-CD can be used as an active pharmaceutical ingredient in the treatment of various neurological diseases, 65 such as inhibition of intracellular cholesterol accumulation through plasma membrane interactions, which plays a beneficial role in Niemann-Pick type C disease, 20,66 and the promising neuroprotective effect in Alzheimer's disease by cholesterol transport and Aβ clearance. 21,67 While previous research has shown the neuroprotective effects of HP-β-CD in MPTP-induced PD mouse models by increasing the autophagy, 22 further independent controlled trials are required to verify the neuroprotective effects of HP-β-CD in PD animal models.…”

Alterations in Proteostasis Mechanisms in Niemann–Pick Type C Disease

“…One treatment that aims to regulate cholesterol levels by improving the solubility of hydrophobic compounds is 2-hydroxypropyl-β-cyclodextrin (HPβCD). However, this drug has been observed to lower cholesterol levels in a model of neuronal NPC cells but did not normalize the levels of NPC1 and [81]. In addition, adverse events such as hearing loss, fever, and meningitis have been reported in some patients [82].…”

Synthetic Receptors for Early Detection and Treatment of Cancer

“…What makes this compound especially important is that it acts against a range of leukemias including acute myeloid leukemia, acute lymphoblastic leukemia and chronic myeloid leukemia as well as inhibiting hypoxia-adapted cells. The compound has also been approved for treating a lysosomal lipid storage disorder, Niemann-Pick Type C disease [87].…”