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Proteomic analysis of the cerebrospinal fluid (CSF) collected from the three oldest patients and three controls (age 14–17 years), performed by a label-free protein quantification approach using nano-ultra-high performance liquid chromatography/nano-electrospray mass spectrometry (nano-UPLC/nano-ESI-MS). a Analysis of the data was performed with the MaxQuant and Perseus quantitative proteomics softwares and illustrated by a volcano plot. Label-free quantitation is shown for b reelin, c Calbindin, d cerebellin-3, and e cerebellin-1, f apolipoprotein B100, and g clusterin in the three oldest A-T patients compared to their controls
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Autosomal recessive ataxia telangiectasia (A-T) is characterized by radiosensitivity, immunodeficiency, and cerebellar neurodegeneration. A-T is caused by inactivating mutations in the ataxia telangiectasiamutated (ATM) gene, a serine-threonine protein kinase involved in DNA damage response and excitatory neurotransmission. The selective vulnerabil...
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... Similarly, the injection of Reelin protein into the cerebellum of reeler mice ameliorated motor function [82]. In addition, decreased levels of Reelin, together with early deficits in Reelin signaling, have been evidenced in the cerebrospinal fluid of patients with Ataxia-telangiectasia (A-T), and a decreased expression of the Reelin receptor was shown in a mouse model of A-T (Atm-/-) [83]. A-T is a rare, inherited debilitating disorder that affects the nervous, immune, and other bodily systems and is characterized by progressive ataxia beginning in early childhood, with difficulty walking, problems with balance and hand coordination, as well as neuropathy. ...
Reelin is an extracellular matrix glycoprotein involved in neuronal migration during embryonic brain development and synaptic plasticity in the adult brain. The role of Reelin in the developing central nervous system has been extensively characterized. Indeed, a loss of Reelin or a disruption in its signaling cascade leads to neurodevelopmental defects and is associated with ataxia, intellectual disability, autism, and several psychiatric disorders. In the adult brain, Reelin is critically involved in neurogenesis and synaptic plasticity. Reelin’s signaling potentiates glutamatergic and GABAergic neurotransmission, induces synaptic maturation, and increases AMPA and NMDA receptor subunits’ expression and activity. As a result, there is a growing literature reporting that a loss of function and/or reduction of Reelin is implicated in numerous neurodegenerative diseases. The present review summarizes the current state of the literature regarding the implication of Reelin and Reelin-mediated signaling during aging and neurodegenerative disorders, highlighting Reelin as a possible target in the prevention or treatment of progressive neurodegeneration.
... To ensure data reproducibility among different organisms in this strongly affected tissue at advanced age, we compared our ATM-null mouse cerebellar transcriptome profile at age 12 months with a published [66] proteome survey of A-T patient cerebellar post-mortem samples (although distortions by altered tissue composition at end-stage will generate artefacts, and mass-spectrometry will detect maximally some 10,000 among all existing proteins), annotating the consistent findings in Table S1. The comparison of our 12-monthold ATM-null mouse cerebellar transcriptome profile with previous A-T patient cerebrospinal fluid proteome data [67] revealed parallel reductions for Reln, Fat2, Omd, Cntn6 (down) and C4b (up). This transcriptome was then interrogated in the context of known ATM functions and phenotypes, as far as known in the current literature. ...
... In the disease context, the transcriptome showed dysregulated expression with nominal significance for genes responsible for phenotypes of ataxia (compiled according to the Online Mendelian Inheritance of Man database, https://www.ncbi.nlm.nih.gov/omim/). Downregulations were observed for Atm, Itpr1, Syne1, Grid2, Grik2, Fgf14, Rora, Gba2, Reln, in good agreement with a previous proteome study of cerebrospinal fluid from A-T patients [67]; a significant enrichment was detected by the STRING webserver for "abnormal cerebellar granule neuron morphology" (q=0.0014) ...
... for the cluster of ATM, RORA [74] and GRID2 [75] proteins; an enrichment for "postsynapse" (q=0.0182) was detected for ITPR1 [66,67,75,76], SYNE1, GRID2 [75] and GRIK2 [77]; upregulations were observed for the ataxia genes Mme, Ebf3, Vamp1, Ppp2r2b, Svbp, without significant enrichment, but VAMP1 being a vesicle-associated factor like ATM. Significant expression changes existed also for genes responsible for the pathogenesis of telangiectasia (upregulation of Sst, Sstr1, Sstr2, Tac1, Tacr1, Svbp) [78][79][80][81], and for general growth (Sst, Sstr1, Sstr2) [82]. ...
The autosomal recessive disorder Ataxia-Telangiectasia is caused by dysfunction of the stress response protein ATM. In the nucleus of proliferating cells, ATM senses DNA double-strand breaks and coordinates their repair. This role explains T-cell dysfunction and tumor risk. However, it remains unclear whether this function is relevant for postmitotic neurons and underlies the cerebellar atrophy, since ATM is cytoplasmic in postmitotic neurons. Here, we used ATM-null mice that survived early immune deficits by bone-marrow transplantation, and reached initial neurodegeneration stages at 12 months of age. Global cerebellar transcriptomics demonstrated ATM depletion to trigger upregulations in most neurotransmission and neuropeptide systems. Downregulated transcripts were found for the ATM interactome component Usp2, many non-coding RNAs, ataxia genes Itpr1, Grid2, immediate early genes and immunity factors. Allelic splice changes affected prominently neuropeptide machinery, e.g. Oprm1. Validation experiments with stressors were performed in human neuroblastoma cells, where ATM localized only to cytoplasm, similar to brain. Effect confirmation in SH-SY5Y cells occurred after ATM depletion and osmotic stress better than nutrient / oxidative stress, not after ATM kinase inhibition or DNA stressor bleomycin. Overall, we provide pioneer observations from a faithful A-T mouse model, which suggest general changes in synaptic and dense-core vesicle stress adaptation.
... GRIN2B is mainly involved in human brain development, not the immune response. However, Canet-Pons et al. [9] reported that GRIN2B was related to the dysregulation of intracellular cholesterol transport in macrophages based on an animal experiment, and George et al. [10] reported that GRIN2B was expressed on the surface of cells in gastric cancer patients with Helicobacter pylori infections. These findings indicate that GRIN2B may have some relationship to the immune response. ...
... However, no children and adolescents were included; therefore, such biomarkers are unable to predict the development of A-T tissue breakdown. In 2018, the research team in Frankfurt demonstrated CSF ApoB excess and Reelin signaling deficits to reflect the neurodegeneration in A-T [47]. Another study of the German A-T cohort detected an altered albumin ratio and altered Reelin levels in CSF. ...
Ataxia telangiectasia (A-T) is a progressive and life-limiting disease associated with cerebellar ataxia due to progressive cerebellar degeneration. In addition to ataxia, which is described in detail, the presence of chorea, dystonia, oculomotor apraxia, athetosis, parkinsonism, and myoclonia are typical manifestations of the disease. The study aimed to evaluate the specificity and sensitivity of neurofilament light chain (NfL) as a biomarker of neurodegeneration in relation to SARA score. In this prospective trial, one visit of 42 A-T patients aged 1.3–25.6 years (mean 11.6 ± 7.3 years) was performed, in which NfL was determined from serum by ELISA. Additionally, a neurological examination of the patients was performed. Blood was collected from 19 healthy volunteers ≥ 12 years of age. We found significantly increased levels of NfL in patients with A-T compared to healthy controls (21.5 ± 3.6 pg/mL vs. 9.3 ± 0.49 pg/mL, p ≤ 0.01). There was a significant correlation of NfL with age, AFP, and SARA. NfL is a new potential progression biomarker in blood for neurodegeneration in A-T which increases with age.
... Evaluating the >2.4-fold changes in Table 1 further, several calciumbinding factors with selective expression in different neuron populations seemed relevant and were highlighted in beige color: The progressive decrease of Calb2 mRNA encoding Calretinin in cerebellum mirrors specifically the GLUergic parallel fibers, whereas the depletion of Parvalbumin (encoded by the Parva gene) is a marker of GABAergic Purkinje neurons. Both are lost from ALS spinal cord tissue (Canet-Pons et al., 2018;Hayashi et al., 2013;Maskey et al., 2010;Wishart et al., 2012). The parallel reduction of Clgn mRNA encoding the endoplasmic reticulum chaperone Calmegin represents cerebellar deep neurons and spinal motor neurons, according to the Allen mouse spinal cord in-situ hybridization data. ...
Large polyglutamine expansions in Ataxin-2 (ATXN2) cause multi-system nervous atrophy in Spinocerebellar Ataxia type 2 (SCA2). Intermediate size expansions carry a risk for selective motor neuron degeneration, known as Amyotrophic Lateral Sclerosis (ALS). Conversely, the depletion of ATXN2 prevents disease progression in ALS. Although ATXN2 interacts directly with RNA, and in ALS pathogenesis there is a crucial role of RNA toxicity, the affected functional pathways remain ill defined. Here, we examined an authentic SCA2 mouse model with Atxn2-CAG100-KnockIn for a first definition of molecular mechanisms in spinal cord pathology. Neurophysiology of lower limbs detected sensory neuropathy rather than motor denervation. Triple immunofluorescence demonstrated cytosolic ATXN2 aggregates sequestrating TDP43 and TIA1 from the nucleus. In immunoblots, this was accompanied by elevated CASP3, RIPK1 and PQBP1 abundance. RT-qPCR showed increase of Grn, Tlr7 and Rnaset2 mRNA versus Eif5a2, Dcp2, Uhmk1 and Kif5a decrease. These SCA2 findings overlap well with known ALS features. Similar to other ataxias and dystonias, decreased mRNA levels for Unc80, Tacr1, Gnal, Ano3, Kcna2, Elovl5 and Cdr1 contrasted with Gpnmb increase. Preterminal stage tissue showed strongly activated microglia containing ATXN2 aggregates, with parallel astrogliosis. Global transcriptome profiles from stages of incipient motor deficit versus preterminal age identified molecules with progressive downregulation, where a cluster of cholesterol biosynthesis enzymes including Dhcr24, Msmo1, Idi1 and Hmgcs1 was prominent. Gas chromatography demonstrated a massive loss of crucial cholesterol precursor metabolites. Overall, the ATXN2 protein aggregation process affects diverse subcellular compartments, in particular stress granules, endoplasmic reticulum and receptor tyrosine kinase signaling. These findings identify new targets and potential biomarkers for neuroprotective therapies.
... The neuropathological changes underlying the cerebellar motor findings in A-T show a macroscopically visible cerebellar atrophy with a thinning of cerebellar hemispheric foliae and cerebellar vermis, which result from a reduction in Purkinje cells, granular cells and basket cells [6,16,17]. In addition, a recent study from our group showed that ApoB excess and reelin signaling deficits reflect the neurodegeneration in A-T in a sensitive and specific way [18]. Imaging studies repeatedly demonstrated cerebellar atrophy in all patients with classic A-T [19,20]. ...
... Thus, there is an unmet need of new surrogate markers of neurodegeneration and disease progression. So far, only two studies (one from our group) used cerebrospinal fluid (CSF) to search for potential biomarker candidates of neural atrophy in A-T [18,23]. Both studies, the one in adult and our recent study in children, applied a highthroughput LC/MS-based label-free protein quantification technology to characterize proteins in CSF samples and to identify differentially expressed proteins, which can serve as biomarker candidates. ...
... There were no significant differences in cell count, lactate, and glucose, whereas total protein and albumin was significantly (p = 0.002) elevated in CSF compared with healthy controls as shown in Table 1. Interestingly, total protein and albumin were significantly correlated to age as recently described [18]. ...
Ataxia telangiectasia (A-T) is a devastating multi-system disorder characterized by progressive cerebellar ataxia and immunodeficiency. The neurological decline may be caused by multiple factors of which ongoing inflammation and oxidative stress may play a dominant role. The objective of the present investigation was to determine cerebrospinal fluid (CSF) proteins and possible low-grade inflammation and its relation to age and neurological deterioration. In the present study, we investigated 15 patients with A-T from 2 to 16 years. Our investigation included blood and CSF tests, clinical neurological examination, A-T score, and MRI findings. The albumin ratio (AR) was analyzed to determine the blood–brain-barrier function. In addition, inflammatory cytokines (IL-1α, IL-6, IL-8, IL-12 p40, IL-17A, IFN-γ, TNF-α) were measured by the multiplex cytometric bead array. We compared the results with those from an age-matched control group. Three of the A-T patients were analyzed separately (one after resection of a cerebral meningioma, one after radiation and chemotherapy due to leukemia, one after stem cell transplantation). Patient had significantly more moderate and severe side effects due to CSF puncture (vomiting, headache, need for anti-emetic drugs) compared with healthy controls. Total protein, albumin, and the AR increased with age indicating a disturbed blood barrier function in older children. There were no differences for cytokines in serum and CSF with the exception of IL-2, which was significantly higher in controls in serum. The AR is significantly altered in A-T patients, but low-grade inflammation is not detectable in serum and CSF.
... In addition, the expression of ITPR1 was reduced in cerebellar neurons derived from A-T patient specific induced pluripotent stem cells, compared with that of normal controls (56). The analyses of cerebrospinal fluid from A-T patients and cerebellar transcripts from Atm −/− animals suggested calcium homeostasis is one of affected physiological conditions in the ATM null cerebellum (57). Furthermore, it has been proposed that an impairment of calcium homeostasis may be predegenerative lesions to Purkinje cell degeneration in A-T patients, as progressive calcium deficits were observed in the Purkinje cells of another Atm null animal model (58). ...
Genomic instability resulting from defective DNA damage responses or repair causes several abnormalities, including progressive cerebellar ataxia, for which the molecular mechanisms are not well understood. Here, we report a new murine model of cerebellar ataxia resulting from concomitant inactivation of POLB and ATM. POLB is one of key enzymes for the repair of damaged or chemically modified bases, including methylated cytosine, but selective inactivation of Polb during neurogenesis affects only a subpopulation of cortical interneurons despite the accumulation of DNA damage throughout the brain. However, dual inactivation of Polb and Atm resulted in ataxia without significant neuropathological defects in the cerebellum. ATM is a protein kinase that responds to DNA strand breaks, and mutations in ATM are responsible for Ataxia Telangiectasia, which is characterized by progressive cerebellar ataxia. In the cerebella of mice deficient for both Polb and Atm, the most downregulated gene was Itpr1, likely because of misregulated DNA methylation cycle. ITPR1 is known to mediate calcium homeostasis, and ITPR1 mutations result in genetic diseases with cerebellar ataxia. Our data suggest that dysregulation of ITPR1 in the cerebellum could be one of contributing factors to progressive ataxia observed in human genomic instability syndromes.
The autosomal recessive disorder Ataxia-Telangiectasia is caused by a dysfunction of the stress response protein, ATM. In the nucleus of proliferating cells, ATM senses DNA double-strand breaks and coordinates their repair. This role explains T-cell dysfunction and tumour risk. However, it remains unclear whether this function is relevant for postmitotic neurons and underlies cerebellar atrophy, since ATM is cytoplasmic in postmitotic neurons. Here, we used ATM-null mice that survived early immune deficits via bone-marrow transplantation, and that reached initial neurodegeneration stages at 12 months of age. Global cerebellar transcriptomics demonstrated that ATM depletion triggered upregulations in most neurotransmission and neuropeptide systems. Downregulated transcripts were found for the ATM interactome component Usp2, many non-coding RNAs, ataxia genes Itpr1, Grid2, immediate early genes and immunity factors. Allelic splice changes affected prominently the neuropeptide machinery, e.g., Oprm1. Validation experiments with stressors were performed in human neuroblastoma cells, where ATM was localised only to cytoplasm, similar to the brain. Effect confirmation in SH-SY5Y cells occurred after ATM depletion and osmotic stress better than nutrient/oxidative stress, but not after ATM kinase inhibition or DNA stressor bleomycin. Overall, we provide pioneer observations from a faithful A-T mouse model, which suggest general changes in synaptic and dense-core vesicle stress adaptation.
Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is one of the most severe global pandemic due to its high pathogenicity and death rate starting from the end of 2019. Though there are some vaccines available against SAER-CoV-2 infections, we are worried about their effectiveness, due to its unstable sequence patterns. Therefore, beside vaccines, globally effective supporting drugs are also required for the treatment against SARS-CoV-2 infection. To explore commonly effective repurposable drugs for the treatment against different variants of coronavirus infections, in this article, an attempt was made to explore host genomic biomarkers guided repurposable drugs for SARS-CoV-1 infections and their validation with SARS-CoV-2 infections by using the integrated bioinformatics approaches. At first, we identified 138 differentially expressed genes (DEGs) between SARS-CoV-1 infected and control samples by analyzing high throughput gene-expression profiles to select drug target key receptors. Then we identified top-ranked 11 key DEGs (SMAD4, GSK3B, SIRT1, ATM, RIPK1, PRKACB, MED17, CCT2, BIRC3, ETS1 and TXN) as hub genes (HubGs) by protein-protein interaction (PPI) network analysis of DEGs highlighting their functions, pathways, regulators and linkage with other disease risks that may influence SARS-CoV-1 infections. The DEGs-set enrichment analysis significantly detected some crucial biological processes (immune response, regulation of angiogenesis, apoptotic process, cytokine production and programmed cell death, response to hypoxia and oxidative stress), molecular functions (transcription factor binding and oxidoreductase activity) and pathways (transcriptional mis-regulation in cancer, pathways in cancer, chemokine signaling pathway) that are associated with SARS-CoV-1 infections as well as SARS-CoV-2 infections by involving HubGs. The gene regulatory network (GRN) analysis detected some transcription factors (FOXC1, GATA2, YY1, FOXL1, TP53 and SRF) and micro-RNAs (hsa-mir-92a-3p, hsa-mir-155-5p, hsa-mir-106b-5p, hsa-mir-34a-5p and hsa-mir-19b-3p) as the key transcriptional and post- transcriptional regulators of HubGs, respectively. We also detected some chemicals (Valproic Acid, Cyclosporine, Copper Sulfate and arsenic trioxide) that may regulates HubGs. The disease-HubGs interaction analysis showed that our predicted HubGs are also associated with several other diseases including different types of lung diseases. Then we considered 11 HubGs mediated proteins and their regulatory 6 key TFs proteins as the drug target proteins (receptors) and performed their docking analysis with the SARS-CoV-2 3CL protease-guided top listed 90 anti-viral drugs out of 3410. We found Rapamycin, Tacrolimus, Torin-2, Radotinib, Danoprevir, Ivermectin and Daclatasvir as the top-ranked 7 candidate-drugs with respect to our proposed target proteins for the treatment against SARS-CoV-1 infections. Then, we validated these 7 candidate-drugs against the already published top-ranked 11 target proteins associated with SARS-CoV-2 infections by molecular docking simulation and found their significant binding affinity scores with our proposed candidate-drugs. Finally, we validated all of our findings by the literature review. Therefore, the proposed candidate-drugs might play a vital role for the treatment against different variants of SARS-CoV-2 infections with comorbidities, since the proposed HubGs are also associated with several comorbidities.
The role of vitamins in the formation of the immune response, both innate and acquired immunity, is well known. At the same time, deficit of fat-soluble vitamins A, E, D leads to impaired response of the immune system to the infectious invasion and to disorders of immune system functioning. The aim of this article is to analyze the literature data on the impact of fat-soluble vitamins on the function of the human immune system and the possibilities of their use in patients with immunodeficiency. Results. Сurrently, there are enough evidences of the successful use of fat-soluble vitamins in secondary immunodeficiencies. Data on the usage of vitamins A, E, D in the treatment of primary immunodeficiencies are few. However, even reducing of antibiotics and other medicines administration in children with primary immunodeficiency indicates the feasibility of their using. The results of scientific studies on the successful use of vitamins D and A in the treatment of allergic diseases, vitamin D in the prevention and improvement of the treatment of autoimmune and oncological diseases indicate the possibility of their use as adjuvant immunomodulatory therapy in children with primary immunodeficiency. Promising may be the use of vitamins A and E, as powerful antioxidants in patients with primary immunodeficiencies with defects in DNA repair processes. Conclusion. The analysis of the literature data has shown that despite many questions need to be resolved, fat-soluble vitamins A, D, and E, and their analogues can be used in clinical settings to enhance the therapeutic effect in children with immune deficiency.
