Rho kinase inhibition modulates microglia activation and improves survival in a model of amyotrophic lateral sclerosis

Abstract

Disease progression in amyotrophic lateral sclerosis (ALS) is characterized by degeneration of motoneurons (MN) and their axons, but is also influenced by neighboring cells such as astrocytes and microglial cells. The role of microglia in ALS is complex as it switches from an anti-inflammatory and neuroprotective phenotype in early disease to a proinflammatory and neurotoxic phenotype in later stages. Our previous studies in models of neurodegeneration identified rho kinase (ROCK) as a target, which can be manipulated to beneficially influence disease progression. Here, we examined the neuroprotective potential of the ROCK inhibitor Fasudil to target the central pathogenic features of ALS. Application of Fasudil to kainic acid-lesioned primary MN in vitro resulted in a strong prosurvival effect. In vivo, SOD1(G93A) mice benefited from oral treatment with Fasudil showing prolonged survival and improved motor function. These findings were correlated to an improved survival of motor neurons and a pronounced alteration of astroglial and microglial cell infiltration of the spinal cord under Fasudil treatment. Modeling a proinflammatory microglial phenotype by stimulation with LPS in vitro, Fasudil decreased the release of proinflammatory cytokines and chemokines TNFα, Il6, CCL2, CCL3, and CCL5 while CXCL1 release was only transiently suppressed. In sciatic nerve motor axons, neuromuscular junction remodeling processes were increased. In conclusion, we provide preclinical and neurobiological evidence that inhibition of ROCK by the clinically approved small molecule inhibitor Fasudil may be a novel therapeutic approach in ALS combining both neuroprotection and immunomodulation for the cure of this devastating disease.

Modulation of Microglial Activity by Rho-Kinase (ROCK) Inhibition as Therapeutic Strategy in Parkinson’s Disease and Amyotrophic Lateral Sclerosis

“…These phenotypic shifts were associated with a decrease in the release of the pro-inflammatory factors NO, IL-1β, IL-6 and TNF-α and increased the release of the anti-inflammatory factor IL-10 in cultured microglia (Ding et al, 2010). A similar suppression of the pro-inflammatory cytokines was observed in a mouse model after Fasudil-treatment (Tönges et al, 2014). Taken together, there is robust evidence that ROCK activity plays an important role in activation of microglia and the determination of its reactive phenotype.…”

Characterization of Motor Neuron Prostaglandin E2 EP3 Receptor Isoform in a Mouse Model of Amyotrophic Lateral Sclerosis

“…These results are consistent with previous results shown that the number of motor neurons in G93A tended to decrease at the early symptomatic stage, but there was no significant difference in the number of the cells between G93A and WT mice. 17) On the other hand, some groups reported that the number of motor neuron decreased significantly in G93A mice at 15 weeks of age. 18,19) Taken together, these data suggested that the characteristics of the motor neurons depend on each G93A mice even carry the same SOD1 mutations.…”

Современные Возможности Идентификации Новых Молекул-Мишеней Для Фармакологической Коррекции Проницаемости Гематоэнцефалического Барьера

“…Ïîä êîí-òðîëåì àêòèâíîñòè Rac1 íàõîäÿòñÿ ñëåäóþùèå ñîáû-òèÿ, àêòóàëüíûå äëÿ ôóíêöèîíèðîâàíèÿ êëåòîê: 1) ýêñ-ïðåññèÿ HIF-1, îïðåäåëÿþùàÿ õàðàêòåð îòâåòà êëåòîê íà ãèïîêñèþ, ýôôåêòû èíñóëèíà è ìåòàáîëèçì ãëþêî-çû; 2) ýêñïðåññèÿ è âíóòðèêëåòî÷íûé òðàíñïîðò òðàíñïîðòåðà ãëþêîçû GLUT4; 3) èçìåíåíèÿ áåëêîâ öèòîñêåëåòà, â ÷àñòíîñòè, íåîáõîäèìûå äëÿ ñòåëëàöèè àñòðîöèòîâ; 4) ïðîäóêöèÿ ñâîáîäíûõ ðàäèêàëîâ, ñåê-ðåöèÿ ìàòðèêñíûõ ìåòàëëîïðîòåèíàç, âàaeíûõ äëÿ ðå-ìîäåëèðîâàíèÿ âíåêëåòî÷íîãî ìàòðèêñà, ÷òî, â öåëîì, ñîîáùàåò ýòîìó áåëêó ïîòåíöèàëüíî âàaeíóþ ðîëü â ðåãóëÿöèè ôóíêöèîíèðîâàíèÿ êëåòîê ÍÂÅ. Ìîäóëÿ-öèÿ Rac1-çàâèñèìûõ ñèãíàëüíûõ ïóòåé òåñòèðóåòñÿ êàê ìåòîä òåðàïèè ïðè íåéðîäåãåíåðàòèâíûõ çàáîëåâà-íèÿõ [48,71,72]. Rac1 òàêaeå ÿâëÿåòñÿ âàaeíûì ðåãóëÿ-òîðîì ÍÀÄÔÍ-îêñèäàçû â ìåìáðàíàõ êëåòîê, NF-kB-êîíòðîëèðóåìîé òðàíñêðèïöèè ïðîâîñïàëè-òåëüíûõ ãåíîâ, ÷òî ìîaeåò îïðåäåëÿòü åãî çíà÷åíèå â ðåãóëÿöèè àêòèâíîñòè ìèêðîãëèè è ìèãðèðóþùèõ â òêàíü ãîëîâíîãî ìîçãà ïðè âîñïàëåíèè ëåéêîöèòîâ, ïðîõîäÿùèõ ÷åðåç ÃÝÁ [25].…”