Synthesis and Development of a Novel First-in-Class Cofilin Inhibitor for Neuroinflammation in Hemorrhagic Brain Injury

You'll have to make sure you don't have this type of stroke, nothing is known on how to recover from it. Lots of big words here, couldn't figure out what it said.

Synthesis and Development of a Novel First-in-Class Cofilin Inhibitor for Neuroinflammation in Hemorrhagic Brain Injury

 

• Saleh I. Alaqel
, 
• Samkeliso Dlamini
, 
• Daniyah A. Almarghalani
, 
• Arjun Shettigar
, 
• Qasim Alhadidi
, 
• Sinali H. Kodithuwakku
, 
• Creed Stary
, 
• L. M. Viranga Tillekeratne
, and 
• Zahoor A. Shah*

Cite this: ACS Chem. Neurosci. 2022, XXXX, XXX, XXX-XXX

Publication Date:March 18, 2022

https://doi.org/10.1021/acschemneuro.2c00010

© 2022 American Chemical Society

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Abstract

Intracerebral hemorrhage (ICH) is devastating among stroke types with high mortality. To date, not a single therapeutic intervention has been successful. Cofilin plays a critical role in inflammation and cell death. In the current study, we embarked on designing and synthesizing a first-in-class small-molecule inhibitor of cofilin to target secondary complications of ICH, mainly neuroinflammation. A series of compounds were synthesized, and two lead compounds SZ-3 and SK-1-32 were selected for further studies. Neuronal and microglial viabilities were assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay using neuroblastoma (SHSY-5Y) and human microglial (HMC-3) cell lines, respectively. Lipopolysaccharide (LPS)-induced inflammation in HMC-3 cells was used for neurotoxicity assay. Other assays include nitric oxide (NO) by Griess reagent, cofilin inhibition by F-actin depolymerization, migration by scratch wound assay, tumor necrosis factor (TNF-α) by enzyme-linked immunosorbent assay (ELISA), protease-activated receptor-1 (PAR-1) by immunocytochemistry and Western blotting (WB), and protein expression levels of several proteins by WB. SK-1-32 increased neuronal/microglial survival, reduced NO, and prevented neurotoxicity. However, SZ-3 showed no effect on neuronal/microglial survival but prevented microglia from LPS-induced inflammation by decreasing NO and preventing neurotoxicity. Therefore, we selected SZ-3 for further molecular studies, as it showed potent anti-inflammatory activities. SZ-3 decreased cofilin severing activity, and its treatment of LPS-activated HMC-3 cells attenuated microglial activation and suppressed migration and proliferation. HMC-3 cells subjected to thrombin, as an in vitro model for hemorrhagic stroke, and treated with SZ-3 after 3 h showed significantly decreased NO and TNF-α, significantly increased protein expression of phosphocofilin, and decreased PAR-1. In addition, SZ-3-treated SHSY-5Y showed a significant increase in cell viability by significantly reducing nuclear factor-κ B (NF-κB), caspase-3, and high-temperature requirement (HtrA2). Together, our results support the novel idea of targeting cofilin to counter neuroinflammation during secondary injury following ICH.

KEYWORDS:

• cofilin inhibitor
• neuroinflammation
• anti-inflammatory agent
• brain injury
• microglial activation

The cytoskeleton of neurons found to play role in Alzheimer's disease

You will need to know this. What EXACTLY  is your doctor doing to followup and use protocols that prevent you from getting Alzheimers?

YOUR DOCTORS' RESPONSIBILITY!

Your chances of getting dementia.

1. A documented 33% dementia chance post-stroke from an Australian study?   May 2012.

2. Then this study came out and seems to have a range from 17-66%. December 2013.

3. A 20% chance in this research.   July 2013.

4. Dementia Risk Doubled in Patients Following Stroke September 2018 

5. Parkinson’s Disease May Have Link to Stroke March 2017

 

The cytoskeleton of neurons found to play role in Alzheimer's disease

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Reviewed by Alina Shrourou, BScJan 17 2019

A researcher at the UPV/EHU-University of the Basque Country has participated in the study into the impairment of the dynamics of spines that receive information from other neurons
A researcher at the UPV/EHU has participated in a study describing what it is during the early stages of Alzheimer's that triggers the loss of dynamics and subsequent impairment of the dendritic spines, the compartments of the neurons responsible for receiving nerve impulses from other neurons. The role played by the actin cytoskeleton of these compartments and how it responds in the presence of beta-amyloid peptides, the component most commonly associated with Alzheimer's, have been described.
José Martínez-Hernández, an Ikerbasque researcher in the Neuronal Ubiquitin Pathways group in the Department of Biochemistry and Molecular Biology of the UPV/EHU's Faculty of Science and Technology, has participated in a study by the Grenoble Institut des Neurosciences which describes the relationship existing between the presence of beta amyloid peptides, known to be the components of the plaques that build up in the brains of people affected by Alzheimer's, and the rapid fall in the dynamism of the actin cytoskeleton of the dendritic spines, the neural area responsible for receiving information coming from other neurons by means of nerve impulses. This reduced dynamics prevents information from being transmitted as it should, and this ultimately triggers the loss of spines and, therefore, the synaptic capacity of the neurons.
The cytoskeleton is a three-dimensional network of proteins that provides cells with internal support, organizes their structures and intervenes in processes such as intracellular transport or traffic. One of the components of the cytoskeleton are the actin filaments which, as Dr Martínez describes, "are anchored but are constantly moving as if they were an escalator; a protein known as cofilin 1 undertakes to sever the filaments and separate the actin units, a task that keeps the dynamics active".
However, if the cofilin 1 has become phosphorylated, in other words, if a phosphorous atom is added to it, this protein moves into an inactive state, it ceases to carry out its function and at the same time prevents neural activity from taking place correctly. "In our study we analyzed samples from human brains with Alzheimer's as well as animal models of this disease, and in them we saw that the inactive form of cofilin 1 is present in higher quantities than in healthy neurons."

Relationship between beta amyloid peptides, phosphorylated cofilin 1 and the ROCK enzyme

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In neuronal cultures they saw that exposure to beta amyloid peptides, the main component of the plaques or deposits that build up in the brains of individuals with Alzheimer's, leads to an increase in phosphorylated cofilin 1, and therefore causes the actin filaments to become too stabilized; the latter lose dynamism and alter the functioning of the dendritic spines. "What is more, the beta amyloid peptides lead to fewer spines in the long term; when they cease to be functional, they are gradually lost over time," highlighted the researcher.
One of the pathways of cofilin 1 phosphorylation is a kinase known as ROCK, a kind of enzyme that modifies other molecules by means of phosphorylation, sometimes activating them and other times deactivating them. In the study they wanted to see whether  ROCK enzyme, a drug used in clinical practice, the function of which is to inhibit the action of the ROCK enzyme, reversed the effect observed in the actin filaments and "we saw that it did. We have not come up with an action mechanism, but we confirmed that the inhibition of the phosphorylation pathway of cofilin 1 prevents exposure to beta amyloid peptides from causing the deactivation of the protein, and the consequent effect on the cytoskeleton of the dendritic spines," said Martínez.
"Our results support the idea that the damage caused by the beta amyloid peptides on the level of the dendritic spines during the early stages of the disease may be prevented by modulating ROCK and cofilin 1, and that, therefore, research needs to be conducted into medications that specifically stop that phosphorylation of cof1 in neurons, so that future medical treatments to combat Alzheimer's disease can be produced," concluded Dr Martínez. ­

Source:

https://www.ehu.eus/en/-/neuronen-zitoeskeletoak-alzheimerren-gaitzean-parte-hartzen-duela-ikusi-dute

Peptide regulation of cofilin activity in the CNS: A novel therapeutic approach for treatment of multiple neurological disorders

Sounds pretty good. You will need to hire your own researcher to follow this up and see where is might be useful for humans.

Peptide regulation of cofilin activity in the CNS: A novel therapeutic approach for treatment of multiple neurological disorders

• Alisa E. Shaw,
• James R. Bamburg^,

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http://dx.doi.org/10.1016/j.pharmthera.2017.02.031

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---

Abstract

Cofilin is a ubiquitous protein which cooperates with many other actin-binding proteins in regulating actin dynamics. Cofilin has essential functions in nervous system development including neuritogenesis, neurite elongation, growth cone pathfinding, dendritic spine formation, and the regulation of neurotransmission and spine function, components of synaptic plasticity essential for learning and memory. Cofilin's phosphoregulation is a downstream target of many transmembrane signaling processes, and its misregulation in neurons has been linked in rodent models to many different neurodegenerative and neurological disorders including Alzheimer disease (AD), aggression due to neonatal isolation, autism, manic/bipolar disorder, and sleep deprivation. Cognitive and behavioral deficits of these rodent models have been largely abrogated by modulation of cofilin activity using viral-mediated, genetic, and/or small molecule or peptide therapeutic approaches. Neuropathic pain in rats from sciatic nerve compression has also been reduced by modulating the cofilin pathway within neurons of the dorsal root ganglia. Neuroinflammation, which occurs following cerebral ischemia/reperfusion, but which also accompanies many other neurodegenerative syndromes, is markedly reduced by peptides targeting specific chemokine receptors, which also modulate cofilin activity. Thus, peptide therapeutics offer potential for cost-effective treatment of a wide variety of neurological disorders. Here we discuss some recent results from rodent models using therapeutic peptides with a surprising ability to cross the rodent blood brain barrier and alter cofilin activity in brain. We also offer suggestions as to how neuronal-specific cofilin regulation might be achieved.

Abbreviations

• Aβ, β-amyloid peptide produced from amyloid precursor protein in AD;
• AD, Alzheimer disease;
• ADF, actin depolymerizing factor;
• AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid;
• APP, amyloid precursor protein;
• APPsw, human APP with Swedish mutation;
• Arp2/3 complex, actin-related proteins 2/3 complex;
• CCR2 and CCR5, G-protein coupled chemokine receptors;
• CIN, chronophin, a cofilin phosphatase (aka pyridoxal-5′-phosphate phosphatase);
• CNS, central nervous system;
• CPP, cell penetrating peptide;
• CTD, C-terminal domain;
• DAPTA, d-ala peptide T-amide (d-ala-STTTNYT-amide);
• DOCK, dedicator of cytokinesis;
• DS, Down syndrome;
• F-actin, filamentous actin;
• G-actin, globular actin (monomeric);
• HIV, human immunodeficiency virus;
• LIMK, a cofilin kinase with a LIM domain;
• LTD, long-term depression;
• LTP, long-term potentiation;
• nArgBP2, neural Abelson-related gene-binding protein 2, product of the Sorbs2 gene NLG1: neuroligin 1;
• NMDA, N-methyl-d-aspartate;
• NOX, NADPH oxidase;
• Pak, p21-activated kinase;
• PDZ, a structural domain first identified in the proteins PSD-95, Dlg1 and ZO-1;
• PKCα, protein kinase C alpha;
• PP2B, protein phosphatase 2B (aka calcineurin);
• PrP^C, cellular prion protein;
• PS1₂, presenilin 1 with M146L and L286V mutations;
• PS1ΔE9, human presenilin 1 missing exon 9;
• pS3 peptide, the S3 peptide phosphorylated on serine 3;
• RanBP9, Ran binding protein 9;
• RAP-310, an all D-amino acid version of DAPTA;
• ROCK, Rho kinase;
• ROS, reactive oxygen species;
• S3 peptide, The N-terminal 16 amino acids of mammalian cofilin;
• Shank3, SH3 and multiple Ankyrin repeat domains 3, also known as ProSAP2;
• SPAR, spine-associated Rap GTPase-activating protein;
• Srv2, named for suppressor of RAS2-val19 allele 2. Also known as cyclase associated protein (CAP or Srv2/CAP);
• SSH, slingshot phosphatase;
• TAT, transactivator of transcription from the human immunodeficiency virus;
• Tβ4, thymosin β4;
• Tpm, tropomyosin;
• WAVE1, Wiskott-Aldrich Syndrome protein-family verprolin homologous protein 1

Keywords

• Dendritic spines;
• Cofilin phosphoregulation;
• Cognitive disorders;
• Psychiatric disorders;
• Neuropathic pain;
• Sleep deprivation;
• Rodent models

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Corresponding author at: Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523-1870, United States.