Dual effect of TAT functionalized DHAH lipid nanoparticles with neurotrophic factors in human BBB and microglia cultures

Abstract

Background Neurodegenerative diseases (NDs) are an accelerating global health problem. Nevertheless, the stronghold of the brain- the blood–brain barrier (BBB) prevents drug penetrance and dwindles effective treatments. Therefore, it is crucial to identify Trojan horse-like drug carriers that can effectively cross the blood–brain barrier and reach the brain tissue. We have previously developed polyunsaturated fatty acids (PUFA)-based nanostructured lipid carriers (NLC), namely DHAH-NLC. These carriers are modulated with BBB-permeating compounds such as chitosan (CS) and trans-activating transcriptional activator (TAT) from HIV-1 that can entrap neurotrophic factors (NTF) serving as nanocarriers for NDs treatment. Moreover, microglia are suggested as a key causative factor of the undergoing neuroinflammation of NDs. In this work, we used in vitro models to investigate whether DHAH-NLCs can enter the brain via the BBB and investigate the therapeutic effect of NTF-containing DHAH-NLC and DHAH-NLC itself on lipopolysaccharide-challenged microglia. Methods We employed human induced pluripotent stem cell-derived brain microvascular endothelial cells (BMECs) to capitalize on the in vivo-like TEER of this BBB model and quantitatively assessed the permeability of DHAH-NLCs. We also used the HMC3 microglia cell line to assess the therapeutic effect of NTF-containing DHAH-NLC upon LPS challenge. Results TAT-functionalized DHAH-NLCs successfully crossed the in vitro BBB model, which exhibited high transendothelial electrical resistance (TEER) values (≈3000 Ω*cm2). Specifically, the TAT-functionalized DHAH-NLCs showed a permeability of up to 0.4% of the dose. Furthermore, using human microglia (HMC3), we demonstrate that DHAH-NLCs successfully counteracted the inflammatory response in our cultures after LPS challenge. Moreover, the encapsulation of glial cell-derived neurotrophic factor (GNDF)-containing DHAH-NLCs (DHAH-NLC-GNDF) activated the Nrf2/HO-1 pathway, suggesting the triggering of the endogenous anti-oxidative system present in microglia. Conclusions Overall, this work shows that the TAT-functionalized DHAH-NLCs can cross the BBB, modulate immune responses, and serve as cargo carriers for growth factors; thus, constituting an attractive and promising novel drug delivery approach for the transport of therapeutics through the BBB into the brain.

Nanomedical research and development in Spain: improving the treatment of diseases from the nanoscale

“…They quantitatively assessed the permeability of DHAH-NLCs in endothelial cells (BMECs). Successfully, they reported that TAT-functionalized DHAH-NLCs successfully crossed the BBB in a cellular model (Hernando et al, 2022).…”

Nanomaterial payload delivery to central nervous system glia for neural protection and repair

“…NPs, including polymer, dendrimer, lipid, extracellular vesicles, and inorganic nanoparticles, have been designed to deliver APIs to microglia in many different CNS disorders and studied in various models, including autism spectrum disorder, cerebral palsy, neuropathic pain, SCI, Alzheimer’s, experimental autoimmune encephalomyelitis (EAE), TBI, retinal degeneration, Rett syndrome, and stroke ( Iezzi et al, 2012 ; Sharma et al, 2017 , 2020 ; Ganbold et al, 2020 ; Khoury et al, 2020 ; Liao et al, 2020 ; Gao et al, 2021 ; Kim et al, 2021 ; Liu et al, 2021 ; He et al, 2022 ; Sepasi et al, 2023 ) ( Table 1 ). API payloads range from small molecule drugs to proteins, peptides, and siRNA ( Lee et al, 2021 ; Liu et al, 2021 ; Hernando et al, 2022 ; Zhang M. et al, 2023 ). Most NP API therapies aim to modulate the microglial inflammatory response, polarizing microglia towards the more neuronally protective M2 phenotype to alleviate the inflammatory response and improve functional outcomes ( Papa et al, 2013 , 2016 ; Lu et al, 2014 ; Nance et al, 2015 , 2017 ; Saxena et al, 2015 ; Kim et al, 2017 ; Wang Y. et al, 2018 ; Ellert-Miklaszewska et al, 2019 ; Cahalane et al, 2020 ; Cho et al, 2021 ; Xiao et al, 2021 ; Baghbanbashi et al, 2022 ; Ganbold et al, 2022 ; Guo et al, 2022 ; Hollinger et al, 2022 ; Shin et al, 2022 ; Ishida et al, 2023 ; Kalashnikova et al, 2023 ; Pu et al, 2023 ).…”