Амилоидные нити
Dec. 17th, 2021 11:02 pm![[personal profile]](https://www.dreamwidth.org/img/silk/identity/user.png){kind=small}
chuka_lisСогласно гипотезе шведских исследователей, и проведенных ими экспериментов ин витро, шипиковый белок коронавируса, целый или частично "порезанный" протеазами, может формировать амилоидные нити в тканях и крови, включая с вовлечением других белков организма. Это может пояснять, дополнительно, его "гиперкоагуляционные" способности, уже отмеченные в других статьях. И, так же, ряд патологий, вызываемых что инфекцией, что даже и вакцинацией.
SARS-CoV-2 infection is associated with a surprising number of morbidities. Uncanny similarities with amyloid-disease associated blood coagulation and fibrinolytic disturbances together with neurologic and cardiac problems led us to investigate the amyloidogenicity of the SARS-CoV-2 Spike protein (S-protein). Amyloid fibril assays of peptide library mixtures and theoretical predictions identified seven amyloidogenic sequences within the S-protein. All seven peptides in isolation formed aggregates during incubation at 37°C. Three 20-amino acid long synthetic Spike peptides (sequence 191-210, 599-618, 1165-1184) fulfilled three amyloid fibril criteria: nucleation dependent polymerization kinetics by ThT, Congo red positivity and ultrastructural fibrillar morphology. Full-length folded S-protein did not form amyloid fibrils, but amyloid-like fibrils with evident branching were formed during 24 hours of S-protein co-incubation with the protease neutrophil elastase (NE) in vitro. NE efficiently cleaved S-protein rendering exposure of amyloidogenic segments and accumulation of the peptide 193-202, part of the most amyloidogenic synthetic Spike peptide. NE is overexpressed at inflamed sites of viral infection and at vaccine injection sites. Our data propose a molecular mechanism for amyloidogenesis of SARS-CoV-2 S-protein in humans facilitated by endoproteolysis. The potential implications of S-protein amyloidogenesis in COVID-19 disease associated pathogenesis and consequences following S-protein based vaccines should be addressed in understanding the disease, long COVID-19, and vaccine side effects.
What ar etheimplicationsofourfindings?COVID‐19pathogenesisismultifactorialandcomplex[18].SevereCOVID‐19 include acute respiratory distress syndrome (ARDS) from severe innate immunesys‐teminflammatoryre actions resulting in lung damage [19]; Cytokine storm [20]; Heart damage, including inflammation of the heart muscle; Kidney damage; Neurological damage; Damage to the circulatory system resulting in poor blood flow; Long‐COVID symptoms include persistent emotional illness and other mental health conditionsresemblingneurodegenerativediseases[18].It has been proposed that severe inflammatory disease including ARDSin combination withSARS‐CoV‐2protein aggregation migh tinduce systemic AAamyloidosis[21]. Neurotropic colonization and cross‐seeding of S‐protein amyloid fibrils to induce aggregation of human endogenous proteins has been discussed in the context of neurodegeneration [4]. Notably, blood clotting associated with extra‐cellular amyloidoticfibrillar aggregates inthe bloodstream have been reported in affected COVID‐19 patients[ 22]. Hypercoagulation and impaired fibrinolysis were demonstrated inexperimentally S‐pro‐tein spike dblood plasma from healthy donors[22]. Similarly, amyloidos isis associated with cerebralamyloidangiopathy, bloodcoagulationdisruption, fibrinolyticdisturbance[23,24] and FXIIKal‐likrein/ Kininactivation andthromboinflammation [25],suggestingpotentiallinksbetweenamyloido‐genesisofS‐proteinandCOVID‐19phenotypes. Inconclusion, wehereinproposedarathersimplemolecularmechanismforhowSARS‐CoV‐2S‐pro‐tein endoproteolysis by NE can render exceptionall yamyloidogenic S‐peptides such assegment 193‐202,and exposure of multipleamyloidogenic segments in proteolytically nicked S‐protein. It is possible that other amyloidogenic peptides and S‐protein nicked by other proteases could be involved. We found tha tall common coronaviruses infecting humans contain amyloidogenic sequences (SFig.5A). Nonetheless, the magnitude of the diverse COVID‐19 symptoms was not previously reported. The seg‐ment 193‐212 is unique for and SARS‐CoV‐2 (SFig.5B), which in combination with acute inflammation could explai nthe putative COVID‐19 associated amyloidosis. Furthermore,over the course of the year 2021 over 8 billion doses of COVID‐19 vaccines have beenadministeredworldwide.Most doses have provided SARS‐CoV‐2 S‐protein as main antigen. A recent case report describes theserendipitous discoveryof amyloid formationina human patient locally at the site of vaccination and in aproximal lymph node within 24 hours ofthe firstdoseofmRNAvaccinecodingforS‐protein [26]. Neutrophil recruitment and activation at the site of vaccination is expected.The localizedamyloidwasdetectedbytheAβ amyloidPETtracer 18F‐ Florbetaben also known to bind toAL,AA and ATTR cardiac amyloid fibrils.There actantalleged amyloidogenic protein inthe vaccinated patient was not identified. We tested thefluorescentamyloidligandsCN‐PiB (fluorescent benzothia‐zole analogue of Pittsburgh compound B) and DF‐9 (fluorescent stilbene analogue of Florbetaben) and found strong fluorescence response towards Spike19 1fibrilsinvitro (SFig.6),supporting thepossibilitythatvaccineinducedS‐protein derivedamyloiddepositionwasdetectedinthe human PETimagingcasestudy[26]. Ifthehereinproposed mechanismforS‐proteinamyloidformationisassociatedwithreportedcar‐diac,‐ blood,‐ and nervous systemdis orders in certain vaccinated individuals [27] is not known,neither are the long‐term consequences,but is trongly recommended to be investigated in this context.
SARS-CoV-2 infection is associated with a surprising number of morbidities. Uncanny similarities with amyloid-disease associated blood coagulation and fibrinolytic disturbances together with neurologic and cardiac problems led us to investigate the amyloidogenicity of the SARS-CoV-2 Spike protein (S-protein). Amyloid fibril assays of peptide library mixtures and theoretical predictions identified seven amyloidogenic sequences within the S-protein. All seven peptides in isolation formed aggregates during incubation at 37°C. Three 20-amino acid long synthetic Spike peptides (sequence 191-210, 599-618, 1165-1184) fulfilled three amyloid fibril criteria: nucleation dependent polymerization kinetics by ThT, Congo red positivity and ultrastructural fibrillar morphology. Full-length folded S-protein did not form amyloid fibrils, but amyloid-like fibrils with evident branching were formed during 24 hours of S-protein co-incubation with the protease neutrophil elastase (NE) in vitro. NE efficiently cleaved S-protein rendering exposure of amyloidogenic segments and accumulation of the peptide 193-202, part of the most amyloidogenic synthetic Spike peptide. NE is overexpressed at inflamed sites of viral infection and at vaccine injection sites. Our data propose a molecular mechanism for amyloidogenesis of SARS-CoV-2 S-protein in humans facilitated by endoproteolysis. The potential implications of S-protein amyloidogenesis in COVID-19 disease associated pathogenesis and consequences following S-protein based vaccines should be addressed in understanding the disease, long COVID-19, and vaccine side effects.
What ar etheimplicationsofourfindings?COVID‐19pathogenesisismultifactorialandcomplex[18].SevereCOVID‐19 include acute respiratory distress syndrome (ARDS) from severe innate immunesys‐teminflammatoryre actions resulting in lung damage [19]; Cytokine storm [20]; Heart damage, including inflammation of the heart muscle; Kidney damage; Neurological damage; Damage to the circulatory system resulting in poor blood flow; Long‐COVID symptoms include persistent emotional illness and other mental health conditionsresemblingneurodegenerativediseases[18].It has been proposed that severe inflammatory disease including ARDSin combination withSARS‐CoV‐2protein aggregation migh tinduce systemic AAamyloidosis[21]. Neurotropic colonization and cross‐seeding of S‐protein amyloid fibrils to induce aggregation of human endogenous proteins has been discussed in the context of neurodegeneration [4]. Notably, blood clotting associated with extra‐cellular amyloidoticfibrillar aggregates inthe bloodstream have been reported in affected COVID‐19 patients[ 22]. Hypercoagulation and impaired fibrinolysis were demonstrated inexperimentally S‐pro‐tein spike dblood plasma from healthy donors[22]. Similarly, amyloidos isis associated with cerebralamyloidangiopathy, bloodcoagulationdisruption, fibrinolyticdisturbance[23,24] and FXIIKal‐likrein/ Kininactivation andthromboinflammation [25],suggestingpotentiallinksbetweenamyloido‐genesisofS‐proteinandCOVID‐19phenotypes. Inconclusion, wehereinproposedarathersimplemolecularmechanismforhowSARS‐CoV‐2S‐pro‐tein endoproteolysis by NE can render exceptionall yamyloidogenic S‐peptides such assegment 193‐202,and exposure of multipleamyloidogenic segments in proteolytically nicked S‐protein. It is possible that other amyloidogenic peptides and S‐protein nicked by other proteases could be involved. We found tha tall common coronaviruses infecting humans contain amyloidogenic sequences (SFig.5A). Nonetheless, the magnitude of the diverse COVID‐19 symptoms was not previously reported. The seg‐ment 193‐212 is unique for and SARS‐CoV‐2 (SFig.5B), which in combination with acute inflammation could explai nthe putative COVID‐19 associated amyloidosis. Furthermore,over the course of the year 2021 over 8 billion doses of COVID‐19 vaccines have beenadministeredworldwide.Most doses have provided SARS‐CoV‐2 S‐protein as main antigen. A recent case report describes theserendipitous discoveryof amyloid formationina human patient locally at the site of vaccination and in aproximal lymph node within 24 hours ofthe firstdoseofmRNAvaccinecodingforS‐protein [26]. Neutrophil recruitment and activation at the site of vaccination is expected.The localizedamyloidwasdetectedbytheAβ amyloidPETtracer 18F‐ Florbetaben also known to bind toAL,AA and ATTR cardiac amyloid fibrils.There actantalleged amyloidogenic protein inthe vaccinated patient was not identified. We tested thefluorescentamyloidligandsCN‐PiB (fluorescent benzothia‐zole analogue of Pittsburgh compound B) and DF‐9 (fluorescent stilbene analogue of Florbetaben) and found strong fluorescence response towards Spike19 1fibrilsinvitro (SFig.6),supporting thepossibilitythatvaccineinducedS‐protein derivedamyloiddepositionwasdetectedinthe human PETimagingcasestudy[26]. Ifthehereinproposed mechanismforS‐proteinamyloidformationisassociatedwithreportedcar‐diac,‐ blood,‐ and nervous systemdis orders in certain vaccinated individuals [27] is not known,neither are the long‐term consequences,but is trongly recommended to be investigated in this context.
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Date: 2021-12-20 05:24 pm (UTC)no subject
Date: 2021-12-20 05:54 pm (UTC)