Proteomic profiling in cerebral amyloid angiopathy reveals an overlap with CADASIL highlighting accumulation of HTRA1 and its substrates

Zellner, Andreas; Müller, Stephan; Lindner, Barbara; Beaufort, Nathalie; Rozemüller, Annemieke; Arzberger, Thomas; Gassen, Nils C.; Lichtenthaler, Stefan F.; Küster, Bernhard; Haffner, Christof; Dichgans, Martin

doi:10.1186/s40478-021-01303-6

Cited by 22 publications

(35 citation statements)

References 82 publications

(117 reference statements)

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“… 10 This phenomenon indicated that the fragility of the vascular wall may lead to not only thrombosis but also hemorrhage in this disease. In addition, recent studies found that HTRA1 colocalize with NOTCH3 deposits in the brain vessels of patients with CADASIL 26 and Aβ deposits in the capillaries of patients with cerebral amyloid angiopathy, 27 but no deposits of amyloid or granular osmiophilic material were observed in this case and the 2 previously reported cases. It still remains unclear how the HTRA1 mutations eventually cause cerebral small vessel dysfunction and how these hereditary CSVD-related genes interact with each other.…”

Section: Discussionsupporting

confidence: 59%

Heterozygous HTRA1 Mutations Cause Cerebral Small Vessel Diseases

et al. 2022

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Background and ObjectivesCerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is a rare hereditary cerebrovascular disease caused by homozygous or compound heterozygous variations in the high-temperature requirement A serine peptidase 1 (HTRA1) gene. However, several studies in recent years have found that some heterozygousHTRA1mutations also cause cerebral small vessel disease (CSVD). The current study aims to report the novel genotypes, phenotypes, and histopathologic results of 3 pedigrees of CSVD with heterozygousHTRA1mutation.MethodsThree pedigrees of familiar CSVD, including 11 symptomatic patients and 3 asymptomatic carriers, were enrolled. Whole-exome sequencing was conducted in the probands for identifying rare variants, which were then evaluated for pathogenicity according to the American College of Medical Genetics and Genomics guidelines. Sanger sequencing was performed for validation of mutations in the probands and other family members. The protease activity was assayed for the novel mutations. All the participants received detailed clinical and imaging examinations and the corresponding results were concluded. Hematoma evacuation was performed for an intracerebral hemorrhage patient with the p.Q318H mutation, and the postoperative pathology including hematoma and cerebral small vessels were examined.ResultsThree novel heterozygousHTRA1mutations (p.Q318H, p.V279M, and p.R274W) were detected in the 3 pedigrees. The protease activity was largely lost for all the mutations, confirming that they were loss-of-function mutations. The patients in each pedigree presented with typical clinical and imaging features of CVSD, and some of them displayed several new phenotypes including color blindness, hydrocephalus, and multiple arachnoid cysts. In addition, family 1 is the largest pedigree with heterozygousHTRA1mutation so far and includes homozygous twins, displaying some variation in clinical phenotypes. More importantly, pathologic study of a patient with p.Q318H mutation showed hyalinization, luminal stenosis, loss of smooth muscle cells, splitting of the internal elastic lamina, and intramural hemorrhage/dissection-like structures.DiscussionThese findings broaden the mutational and clinical spectrum of heterozygousHTRA1-related CSVD. Pathologic features were similar with the previous heterozygous and homozygous cases. Moreover, clinical heterogeneity was revealed within the largest single family, and the mechanisms of the phenotypic heterogenetic remain unclear. Overall, heterozygous HTRA1-related CSVD should not be simply taken as a mild type of CARASIL as previously considered.

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Section: Discussionsupporting

confidence: 59%

Heterozygous HTRA1 Mutations Cause Cerebral Small Vessel Diseases

et al. 2022

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“…It is interesting to compare our proteomic data with data reported in previous CAA proteomics studies. In two studies, the brain vessels were isolated from CAA frozen brains and used for proteomic analysis, but the COL6A family, which is directly involved in fibrosis, a typical CAA pathology, was not detected or was not increased in CAA vessels [ 10 , 64 ]. CAA vessels are very fragile and the vascular structure is disrupted during the process of vessel isolation.…”

Section: Resultsmentioning

confidence: 99%

Proteomics-based investigation of cerebrovascular molecular mechanisms in cerebral amyloid angiopathy by the FFPE-LMD-PCT-SWATH method

Handa

Sasaki

Takao

et al. 2022

Fluids Barriers CNS

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Background Cerebral amyloid angiopathy (CAA) occurs in 80% of patients with Alzheimer’s disease (AD) and is mainly caused by the abnormal deposition of Aβ in the walls of cerebral blood vessels. Cerebrovascular molecular mechanisms in CAA were investigated by using comprehensive and accurate quantitative proteomics. Methods Concerning the molecular mechanisms specific to CAA, formalin-fixed paraffin-embedded (FFPE) sections were prepared from patients having AD neuropathologic change (ADNC) with severe cortical Aβ vascular deposition (ADNC +/CAA +), and from patients having ADNC without vascular deposition of Aβ (ADNC +/CAA −; so called, AD). Cerebral cortical vessels were isolated from FFPE sections using laser microdissection (LMD), processed by pressure cycling technology (PCT), and applied to SWATH (sequential window acquisition of all theoretical fragment ion spectra) proteomics. Results The protein expression levels of 17 proteins in ADNC +/CAA +/H donors (ADNC +/CAA + donors with highly abundant Aβ in capillaries) were significantly different from those in ADNC +/CAA − and ADNC −/CAA − donors. Furthermore, we identified 56 proteins showing more than a 1.5-fold difference in average expression levels between ADNC +/CAA + and ADNC −/CAA − donors, and were significantly correlated with the levels of Aβ or Collagen alpha-2(VI) chain (COL6A2) (CAA markers) in 11 donors (6 ADNC +/CAA + and 5 ADNC −/CAA −). Over 70% of the 56 proteins showed ADNC +/CAA + specific changes in protein expression. The comparative analysis with brain parenchyma showed that more than 90% of the 56 proteins were vascular-specific pathological changes. A literature-based pathway analysis showed that 42 proteins are associated with fibrosis, oxidative stress and apoptosis. This included the increased expression of Heat shock protein HSP 90-alpha, CD44 antigen and Carbonic anhydrase 1 which are inhibited by potential drugs against CAA. Conclusions The combination of LMD-based isolation of vessels from FFPE sections, PCT-assisted sample processing and SWATH analysis (FFPE-LMD-PCT-SWATH method) revealed for the first time the changes in the expression of many proteins that are involved in fibrosis, ROS production and cell death in ADNC +/CAA + (CAA patients) vessels. The findings reported herein would be useful for developing a better understanding of the pathology of CAA and for promoting the discovery and development of drugs and biomarkers for CAA.

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“…A similar analysis in patients is currently not possible due to the lack of appropriate post mortem tissue. However, proteomic analyses of vessel samples derived from CAA and CADASIL patients revealed an unexpected link to HTRA1 providing further insight into its role in humans (Zellner et al, 2018(Zellner et al, , 2022.…”

Section: High Temperature Requirement Protein Amentioning

confidence: 99%

“…. (Zellner et al, 2018(Zellner et al, , 2022Kato et al, 2021). Evidence for an impairment of HTRA1-mediated proteolytic activity in the cerebrovasculature has so far only been found in patients carrying congenital mutations (Nozaki et al, 2014), thus HTRA1 inactivation via sequestration on a protein level would represent a novel pathomechanism (Figure 1).…”

Section: Mechanisms Of Htra Inactivationmentioning

confidence: 99%

The emerging role of the HTRA1 protease in brain microvascular disease

Haffner¹

2023

Front. Dement.

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Pathologies of the brain microvasculature, often referred to as cerebral small-vessel disease, are important contributors to vascular dementia, the second most common form of dementia in aging societies. In addition to their role in acute ischemic and hemorrhagic stroke, they have emerged as major cause of age-related cognitive decline in asymptomatic individuals. A central histological finding in these pathologies is the disruption of the vessel architecture including thickening of the vessel wall, narrowing of the vessel lumen and massive expansion of the mural extracellular matrix. The underlying molecular mechanisms are largely unknown, but from the investigation of several disease forms with defined etiology, high temperature requirement protein A1 (HTRA1), a secreted serine protease degrading primarily matrisomal substrates, has emerged as critical factor and potential therapeutic target. A genetically induced loss of HTRA1 function in humans is associated with cerebral autosomal-recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL), a rare, hereditary form of brain microvascular disease. Recently, proteomic studies on cerebral amyloid angiopathy (CAA), a common cause of age-related dementia, and cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), the most prevalent monogenic small-vessel disease, have provided evidence for an impairment of HTRA1 activity through sequestration into pathological protein deposits, suggesting an alternative mechanism of HTRA1 inactivation and expanding the range of diseases with HTRA1 involvement. Further investigations of the mechanisms of HTRA1 regulation in the brain microvasculature might spawn novel strategies for the treatment of small-vessel pathologies.

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Proteomic profiling in cerebral amyloid angiopathy reveals an overlap with CADASIL highlighting accumulation of HTRA1 and its substrates

Cited by 22 publications

References 82 publications

Heterozygous HTRA1 Mutations Cause Cerebral Small Vessel Diseases

Heterozygous HTRA1 Mutations Cause Cerebral Small Vessel Diseases

Proteomics-based investigation of cerebrovascular molecular mechanisms in cerebral amyloid angiopathy by the FFPE-LMD-PCT-SWATH method

The emerging role of the HTRA1 protease in brain microvascular disease

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