bims-axbals Biomed News
on Axonal biology and ALS
Issue of 2024–07–07
seventeen papers selected by
TJ Krzystek, ALS Therapy Development Institute
  1. Aberrant evoked calcium signaling and nAChR cluster morphology in a SOD1 D90A hiPSC-derived neuromuscular model.
  2. Retinal cells derived from patients with DRAM2-dependent CORD21 dystrophy exhibit key lysosomal enzyme deficiency and lysosomal content accumulation.
  3. BI1 Activates Autophagy and Mediates TDP43 to Regulate ALS Pathogenesis.
  4. AAV gene replacement therapy for treating MPS IIIC: Facilitating bystander effects via EV-mRNA cargo.
  5. From whence it came: Mitochondrial mRNA leaves, a protein returns.
  6. STAMBP is Required for Long-Term Maintenance of Neural Progenitor Cells Derived from hESCs.
  7. VAMP2 regulates phase separation of α-synuclein.
  8. TDP-43 in nuclear condensates: where, how, and why.
  9. The Role of Alpha-Synuclein in Synucleinopathy: Impact on Lipid Regulation at Mitochondria-ER Membranes.
  10. Diving deep: zebrafish models in motor neuron degeneration research.
  11. Reversibly-bonded microfluidic devices for stable cell culture and rapid, gentle cell extraction.
  12. Cryo-electron tomography reveals the microtubule-bound form of inactive LRRK2.
  13. N6-methyldeoxyadenosine modification difference contributes to homocysteine-induced mitochondrial perturbation in rat hippocampal primary neurons and PC12 cells.
  14. Reduction of Z alpha-1 antitrypsin polymers in human iPSC-hepatocytes and mice by LRRK2 inhibitors.
  15. HaloTag display enables quantitative single-particle characterisation and functionalisation of engineered extracellular vesicles.
  16. A novel TBK1 loss-of-function variant associated with ALS and parkinsonism phenotypes.
  17. Living Biobanks of Organoids: Valuable Resource for Translational Research.
  1. Front Cell Dev Biol. 2024 ;12 1429759
    Aberrant evoked calcium signaling and nAChR cluster morphology in a SOD1 D90A hiPSC-derived neuromuscular model.
    Nathalie Couturier, Sarah Janice Hörner, Elina Nürnberg, Claudio Joazeiro, Mathias Hafner, Rüdiger Rudolf.
      Familial amyotrophic lateral sclerosis (ALS) is a progressive neuromuscular disorder that is due to mutations in one of several target genes, including SOD1. So far, clinical records, rodent studies, and in vitro models have yielded arguments for either a primary motor neuron disease, or a pleiotropic pathogenesis of ALS. While mouse models lack the human origin, in vitro models using human induced pluripotent stem cells (hiPSC) have been recently developed for addressing ALS pathogenesis. In spite of improvements regarding the generation of muscle cells from hiPSC, the degree of maturation of muscle cells resulting from these protocols has remained limited. To fill these shortcomings, we here present a new protocol for an enhanced myotube differentiation from hiPSC with the option of further maturation upon coculture with hiPSC-derived motor neurons. The described model is the first to yield a combination of key myogenic maturation features that are consistent sarcomeric organization in association with complex nAChR clusters in myotubes derived from control hiPSC. In this model, myotubes derived from hiPSC carrying the SOD1 D90A mutation had reduced expression of myogenic markers, lack of sarcomeres, morphologically different nAChR clusters, and an altered nAChR-dependent Ca2+ response compared to control myotubes. Notably, trophic support provided by control hiPSC-derived motor neurons reduced nAChR cluster differences between control and SOD1 D90A myotubes. In summary, a novel hiPSC-derived neuromuscular model yields evidence for both muscle-intrinsic and nerve-dependent aspects of neuromuscular dysfunction in SOD1-based ALS.
    Keywords:  acetylcholine receptors; amyotrophic lateral sclerosis; hiPSC; motor neurons; myogenesis; neuromuscular junction; skeletal muscle cells; stem cells
    DOI:  https://doi.org/10.3389/fcell.2024.1429759
  2. Stem Cell Reports. 2024 Jun 18. pii: S2213-6711(24)00155-3. [Epub ahead of print]
    Retinal cells derived from patients with DRAM2-dependent CORD21 dystrophy exhibit key lysosomal enzyme deficiency and lysosomal content accumulation.
    Rozaliya Tsikandelova, Eldo Galo, Edvinas Cerniauskas, Dean Hallam, Maria Georgiou, Rodrigo Cerna-Chavez, Robert Atkinson, Pavel Palmowski, Florence Burté, Tracey Davies, David H Steel, Martin McKibbin, Jacquelyn Bond, Jennifer Haggarty, Phil Whitfield, Viktor Korolchuk, Lyle Armstrong, Chunbo Yang, Birthe Dorgau, Marzena Kurzawa-Akanbi, Majlinda Lako.
      Biallelic mutations in DRAM2 lead to an autosomal recessive cone-rod dystrophy known as CORD21, which typically presents between the third and sixth decades of life. Although DRAM2 localizes to the lysosomes of photoreceptor and retinal pigment epithelium (RPE) cells, its specific role in retinal degeneration has not been fully elucidated. In this study, we generated and characterized retinal organoids (ROs) and RPE cells from induced pluripotent stem cells (iPSCs) derived from two CORD21 patients. Our investigation revealed that CORD21-ROs and RPE cells exhibit abnormalities in lipid metabolism, defects in autophagic flux, accumulation of aberrant lysosomal content, and reduced lysosomal enzyme activity. We identified potential interactions of DRAM2 with vesicular trafficking proteins, suggesting its involvement in this cellular process. These findings collectively suggest that DRAM2 plays a crucial role in maintaining the integrity of photoreceptors and RPE cells by regulating lysosomal function, autophagy, and potentially vesicular trafficking.
    Keywords:  CORD21; DRAM2; RPE cells; autophagy; lipd metabolism; lysosomes; retinal organoids; vesicular transport
    DOI:  https://doi.org/10.1016/j.stemcr.2024.06.002
  3. Mol Neurobiol. 2024 Jul 02.
    BI1 Activates Autophagy and Mediates TDP43 to Regulate ALS Pathogenesis.
    Yu Wang, Yuxiang Wang, Hanlan Yin, Zixuan Xiao, Zhichao Ren, Xueting Ma, Jingtian Zhang, Xueqi Fu, Fuqiang Zhang, Linlin Zeng.
      Amyotrophic lateral sclerosis (ALS) is the most prevalent motor neuron disease in adults. Currently, there are no known drugs or clinical approaches that have demonstrated efficacy in treating ALS. Mitochondrial function and autophagy have been identified as crucial mechanisms in the development of ALS. While Bax inhibitor 1 (BI1) has been implicated in neurodegenerative diseases, its exact mechanism remains unknown. This study investigates the therapeutic impact of BI1 overexpression on ALS both in vivo and in vitro, revealing its ability to mitigate SOD1G93A-induced apoptosis, nuclear damage, mitochondrial dysfunction, and axonal degeneration of motor neurons. At the same time, BI1 prolongs onset time and lifespan of ALS mice, improves motor function, and alleviates neuronal damage, muscle damage, neuromuscular junction damage among other aspects. The findings indicate that BI1 can inhibit pathological TDP43 morphology and initially stimulate autophagy through interaction with TDP43. This study establishes a solid theoretical foundation for understanding the regulation of autophagy by BI1 and TDP43 while shedding light on the pathogenesis of ALS through their interaction - offering new concepts and targets for clinical implementation and drug development.
    Keywords:  ALS; Apoptosis; Exosome; MAM; Mitochondria; Neurodegenerative disease
    DOI:  https://doi.org/10.1007/s12035-024-04313-2
  4. J Extracell Vesicles. 2024 Jul;13(7): e12464
    AAV gene replacement therapy for treating MPS IIIC: Facilitating bystander effects via EV-mRNA cargo.
    Tierra A Bobo, Michael Robinson, Christopher Tofade, Marina Sokolski-Papkov, Peter Nichols, Stephen Vorobiov, Haiyan Fu.
      MPS IIIC is a lysosomal storage disease caused by mutations in heparan-α-glucosaminide N-acetyltransferase (HGSNAT), for which no treatment is available. Because HGSNAT is a trans-lysosomal-membrane protein, gene therapy for MPS IIIC needs to transduce as many cells as possible for maximal benefits. All cells continuously release extracellular vesicles (EVs) and communicate by exchanging biomolecules via EV trafficking. To address the unmet need, we developed a rAAV-hHGSNATEV vector with an EV-mRNA-packaging signal in the 3'UTR to facilitate bystander effects, and tested it in an in vitro MPS IIIC model. In human MPS IIIC cells, rAAV-hHGSNATEV enhanced HGSNAT mRNA and protein expression, EV-hHGSNAT-mRNA packaging, and cleared GAG storage. Importantly, incubation with EVs led to hHGSNAT protein expression and GAG contents clearance in recipient MPS IIIC cells. Further, rAAV-hHGSNATEV transduction led to the reduction of pathological EVs in MPS IIIC cells to normal levels, suggesting broader therapeutic benefits. These data demonstrate that incorporating the EV-mRNA-packaging signal into a rAAV-hHGSNAT vector enhances EV packaging of hHGSNAT-mRNA, which can be transported to non-transduced cells and translated into functional rHGSNAT protein, facilitating cross-correction of disease pathology. This study supports the therapeutic potential of rAAVEV for MPS IIIC, and broad diseases, without having to transduce every cell.
    Keywords:  EV‐mRNA cargo; adeno‐associated virus (AAV); blood‐brain‐barrier; bystander effects/cross‐correction; lysosomal storage diseases; mucopolysaccharidosis type IIIC (MPS IIIC); rAAV gene therapy
    DOI:  https://doi.org/10.1002/jev2.12464
  5. Cell Metab. 2024 Jul 02. pii: S1550-4131(24)00227-4. [Epub ahead of print]36(7): 1433-1435
    From whence it came: Mitochondrial mRNA leaves, a protein returns.
    Kevin A Janssen, Angela Song.
      Small peptides have previously been reported to be encoded in mitochondrial rRNA and translated by cytosolic ribosomes. In this issue of Cell Metabolism, Hu et al. use mass spectrometry to identify a cytosolically translated protein, encoded instead in mitochondrial mRNA, that is surprisingly targeted back into the mitochondrial matrix.
    DOI:  https://doi.org/10.1016/j.cmet.2024.06.002
  6. Stem Cell Rev Rep. 2024 Jun 29.
    STAMBP is Required for Long-Term Maintenance of Neural Progenitor Cells Derived from hESCs.
    Jitian Zhang, Yanqi Zhang, Yancai Liu, Tiancheng Zhou, Guangjin Pan, Jufang He, Xiaodong Shu.
      Mutations in STAMBP have been well-established to cause congenital human microcephaly-capillary malformation (MIC-CAP) syndrome, a rare genetic disorder characterized by global developmental delay, severe microcephaly, capillary malformations, etc. Previous biochemical investigations and loss-of-function studies in mice have provided insights into the mechanism of STAMBP, however, it remains controversial how STAMBP deficiency leads to malformation of those affected tissues in patients. In this study, we investigated the function and underlying mechanism of STAMBP during neural differentiation of human embryonic stem cells (hESCs). We found that STAMBP is dispensable for the pluripotency maintenance or neural differentiation of hESCs. However, neural progenitor cells (NPCs) derived from STAMBP-deficient hESCs fail to be long-term maintained/expanded in vitro. We identified the anti-apoptotic protein CFLAR is down-regulated in those affected NPCs and ectopic expression of CFLAR rescues NPC defects induced by STAMBP-deficiency. Our study not only provides novel insight into the mechanism of neural defects in STAMBP mutant patients, it also indicates that the death receptor mediated apoptosis is an obstacle for long-term maintenance/expansion of NPCs in vitro thus counteracting this cell death pathway could be beneficial to the generation of NPCs in vitro.
    Keywords:  Apoptosis; CFLAR; MIC-CAP syndrome; STAMBP; hESCs
    DOI:  https://doi.org/10.1007/s12015-024-10751-1
  7. Nat Cell Biol. 2024 Jul 01.
    VAMP2 regulates phase separation of α-synuclein.
    Aishwarya Agarwal, Aswathy Chandran, Farheen Raza, Irina-Maria Ungureanu, Christine Hilcenko, Katherine Stott, Nicholas A Bright, Nobuhiro Morone, Alan J Warren, Janin Lautenschläger.
      α-Synuclein (αSYN), a pivotal synaptic protein implicated in synucleinopathies such as Parkinson's disease and Lewy body dementia, undergoes protein phase separation. We reveal that vesicle-associated membrane protein 2 (VAMP2) orchestrates αSYN phase separation both in vitro and in cells. Electrostatic interactions, specifically mediated by VAMP2 via its juxtamembrane domain and the αSYN C-terminal region, drive phase separation. Condensate formation is specific for R-SNARE VAMP2 and dependent on αSYN lipid membrane binding. Our results delineate a regulatory mechanism for αSYN phase separation in cells. Furthermore, we show that αSYN condensates sequester vesicles and attract complexin-1 and -2, thus supporting a role in synaptic physiology and pathophysiology.
    DOI:  https://doi.org/10.1038/s41556-024-01451-6
  8. Biochem Soc Trans. 2024 Jul 03. pii: BST20231447. [Epub ahead of print]
    TDP-43 in nuclear condensates: where, how, and why.
    Ruaridh Lang, Rachel E Hodgson, Tatyana A Shelkovnikova.
      TDP-43 is an abundant and ubiquitously expressed nuclear protein that becomes dysfunctional in a spectrum of neurodegenerative diseases. TDP-43's ability to phase separate and form/enter biomolecular condensates of varying size and composition is critical for its functionality. Despite the high density of phase-separated assemblies in the nucleus and the nuclear abundance of TDP-43, our understanding of the condensate-TDP-43 relationship in this cellular compartment is only emerging. Recent studies have also suggested that misregulation of nuclear TDP-43 condensation is an early event in the neurodegenerative disease amyotrophic lateral sclerosis. This review aims to draw attention to the nuclear facet of functional and aberrant TDP-43 condensation. We will summarise the current knowledge on how TDP-43 containing nuclear condensates form and function and how their homeostasis is affected in disease.
    Keywords:  ALS; TDP-43; condensate; neurodegeneration; nuclear body; paraspeckle; stress response
    DOI:  https://doi.org/10.1042/BST20231447
  9. bioRxiv. 2024 Jun 17. pii: 2024.06.17.599406. [Epub ahead of print]
    The Role of Alpha-Synuclein in Synucleinopathy: Impact on Lipid Regulation at Mitochondria-ER Membranes.
    Peter A Barbuti, Cristina Guardia-Laguarta, Taekyung Yun, Zena K Chatila, Xena Flowers, Bruno Fr Santos, Simone B Larsen, Nobutaka Hattori, Elizabeth Bradshaw, Ulf Dettmer, Saranna Fanning, Manon Vilas, Hasini Reddy, Andrew F Teich, Rejko Krüger, Estela Area-Gomez, Serge Przedborski.
      The protein alpha-synuclein (αSyn) plays a critical role in the pathogenesis of synucleinopathy, which includes Parkinson's disease and multiple system atrophy, and mounting evidence suggests that lipid dyshomeostasis is a critical phenotype in these neurodegenerative conditions. Previously, we identified that αSyn localizes to mitochondria-associated endoplasmic reticulum membranes (MAMs), temporary functional domains containing proteins that regulate lipid metabolism, including the de novo synthesis of phosphatidylserine. In the present study, we have analyzed the lipid composition of postmortem human samples, focusing on the substantia nigra pars compacta of Parkinson's disease and controls, as well as three less affected brain regions of Parkinson's donors. To further assess synucleinopathy-related lipidome alterations, similar analyses were performed on the striatum of multiple system atrophy cases. Our data show region-and disease-specific changes in the levels of lipid species. Specifically, our data revealed alterations in the levels of specific phosphatidylserine species in brain areas most affected in Parkinson's disease. Some of these alterations, albeit to a lesser degree, are also observed multiples system atrophy. Using induced pluripotent stem cell-derived neurons, we show that αSyn contributes to regulating phosphatidylserine metabolism at MAM domains, and that αSyn dosage parallels the perturbation in phosphatidylserine levels. Our results support the notion that αSyn pathophysiology is linked to the dysregulation of lipid homeostasis, which may contribute to the vulnerability of specific brain regions in synucleinopathy. These findings have significant therapeutic implications.
    Significance Statement: Synucleinopathy is a complex group of neurodegenerative disorders whose causes and underlying mechanisms remain unknown. In this work, we examined synucleinopathy postmortem brain samples and patient-derived neuron models and identified the functional impairment of the mitochondrial-associated endoplasmic reticulum membrane (MAM) domain, which facilitates lipid regulation. The protein alpha-synuclein is associated with synucleinopathy and increasing levels result in the mislocalization of this protein and the disruption of MAM domains, which, in turn, results in lipid and membrane composition alterations. Specifically, we report that increased alpha-synuclein expression impairs the regulation of phosphatidylserine synthase 2 and the levels of phosphatidylserine in cellular membranes from affected cells. Our study offers mechanistic insight tying alpha-synuclein pathology and lipid dysregulation as seminal factors in synucleinopathy, which may have pathogenic and therapeutic implications.
    DOI:  https://doi.org/10.1101/2024.06.17.599406
  10. Front Neurosci. 2024 ;18 1424025
    Diving deep: zebrafish models in motor neuron degeneration research.
    Vranda Garg, Bart R H Geurten.
      In the dynamic landscape of biomedical science, the pursuit of effective treatments for motor neuron disorders like hereditary spastic paraplegia (HSP), amyotrophic lateral sclerosis (ALS), and spinal muscular atrophy (SMA) remains a key priority. Central to this endeavor is the development of robust animal models, with the zebrafish emerging as a prime candidate. Exhibiting embryonic transparency, a swift life cycle, and significant genetic and neuroanatomical congruencies with humans, zebrafish offer substantial potential for research. Despite the difference in locomotion-zebrafish undulate while humans use limbs, the zebrafish presents relevant phenotypic parallels to human motor control disorders, providing valuable insights into neurodegenerative diseases. This review explores the zebrafish's inherent traits and how they facilitate profound insights into the complex behavioral and cellular phenotypes associated with these disorders. Furthermore, we examine recent advancements in high-throughput drug screening using the zebrafish model, a promising avenue for identifying therapeutically potent compounds.
    Keywords:  amyotrophic lateral sclerosis (ALS); drug screening; hereditary spastic paraplegia (HSP); motor neuron disorders; spinal muscular atrophy (SMA); zebrafish
    DOI:  https://doi.org/10.3389/fnins.2024.1424025
  11. Lab Chip. 2024 Jul 01.
    Reversibly-bonded microfluidic devices for stable cell culture and rapid, gentle cell extraction.
    Xiaohan Feng, Zehaoyu Wu, Lily Kwan Wai Cheng, Yang Xiang, Ryohichi Sugimura, Xuyan Lin, Angela Ruohao Wu.
      Microfluidic chips have emerged as significant tools in cell culture due to their capacity for supporting cells to adopt more physiologically relevant morphologies in 3D compared with traditional cell culture in 2D. Currently, irreversible bonding methods, where chips cannot be detached from their substrates without destroying the structure, are commonly used in fabrication, making it challenging to conduct further analysis on cells that have been cultured on-chip. Although some reversible bonding techniques have been developed, they are either restricted to certain materials such as glass, or require complex processing procedures. Here, we demonstrate a simple and reversible polydimethylsiloxane (PDMS)-polystyrene (PS) bonding technique that allows devices to withstand extended operations while pressurized, and supports long-term stable cell cultures. More importantly, it allows rapid and gentle live cell extraction for downstream manipulation and characterization after long-term on-chip culturing, and even further subculturing. Our new approach could greatly facilitate microfluidic chip-based cell and tissue cultures, overcoming current analytical limitations and opening up new avenues for downstream uses of on-chip cultures, including 3D-engineered tissue structures for biomedical applications.
    DOI:  https://doi.org/10.1039/d3lc01019h
  12. bioRxiv. 2024 Jun 20. pii: 2024.06.18.599606. [Epub ahead of print]
    Cryo-electron tomography reveals the microtubule-bound form of inactive LRRK2.
    Siyu Chen, Tamar Basiashvili, Joshua Hutchings, Marta Sanz Murillo, Amalia Villagran Suarez, Jaime Alegrio Louro, Andres E Leschziner, Elizabeth Villa.
      Parkinson's Disease (PD) is the second most common neurodegenerative disorder. Mutations in leucine-rich repeat kinase 2 (LRRK2), a multi-domain protein containing both a kinase and a GTPase, are a leading cause of the familial form of PD. Pathogenic LRRK2 mutations increase LRRK2 kinase activity. While the bulk of LRRK2 is found in the cytosol, the protein associates with membranes where its Rab GTPase substrates are found, and under certain conditions, with microtubules. Integrative structural studies using single-particle cryo-electron microscopy (cryo-EM) and in situ cryo-electron tomography (cryo-ET) have revealed the architecture of microtubule-associated LRRK2 filaments, and that formation of these filaments requires LRRK2's kinase to be in the active-like conformation. However, whether LRRK2 can interact with and form filaments on microtubules in its autoinhibited state, where the kinase domain is in the inactive conformation and the N-terminal LRR domain covers the kinase active site, was not known. Using cryo-ET, we show that full-length LRRK2 can oligomerize on microtubules in its autoinhibited state. Both WT-LRRK2 and PD-linked LRRK2 mutants formed filaments on microtubules. While these filaments are stabilized by the same interfaces seen in the active-LRRK2 filaments, we observed a new interface involving the N-terminal repeats that were disordered in the active-LRRK2 filaments. The helical parameters of the autoinhibited-LRRK2 filaments are different from those reported for the active-LRRK2 filaments. Finally, the autoinhibited-LRRK2 filaments are shorter and less regular, suggesting they are less stable.
    DOI:  https://doi.org/10.1101/2024.06.18.599606
  13. Biochem Pharmacol. 2024 Jul 03. pii: S0006-2952(24)00393-9. [Epub ahead of print] 116410
    N6-methyldeoxyadenosine modification difference contributes to homocysteine-induced mitochondrial perturbation in rat hippocampal primary neurons and PC12 cells.
    Ling Zhang, Fang Xie, Xue Wang, Zhaowei Sun, Hui Hu, Yuhan Wu, Shijia Zhang, Xiaobing Chen, Lingjia Qian, Yun Zhao.
      Elevated homocysteine (Hcy) levels are detrimental to neuronal cells and contribute to cognitive dysfunction in rats. Mitochondria plays a crucial role in cellular energy metabolism. Interestingly, the damaging effects of Hcy in vivo and in vitro conditions exhibit distinct results. Herein, we aimed to investigate the effects of Hcy on mitochondrial function in primary neurons and PC12 cells and explore the underlying mechanisms involved. The metabolic intermediates of Hcy act as methyl donors and play important epigenetic regulatory roles. N6-methyldeoxyadenosine (6 mA) modification, which is enriched in mitochondrial DNA (mtDNA), can be mediated by methyltransferase METTL4. Our study suggested that mitochondrial perturbation caused by Hcy in primary neurons and PC12 cells may be attributable to mtDNA 6 mA modification difference. Hcy could activate the expression of METTL4 within mitochondria to facilitate mtDNA 6 mA status, and repress mtDNA transcription, then result in mitochondrial dysfunction.
    Keywords:  Homocysteine; Mitochondrial oxidative phosphorylation; N-6-deoxyadenine methylation; PC12 cell line; Primary neurons; Targeted prevention
    DOI:  https://doi.org/10.1016/j.bcp.2024.116410
  14. Hepatology. 2024 Jul 02.
    Reduction of Z alpha-1 antitrypsin polymers in human iPSC-hepatocytes and mice by LRRK2 inhibitors.
    Deniz Kent, Soon Seng Ng, Adam M Syanda, Payam Khoshkenar, Riccardo Ronzoni, Chao Zheng Li, Marina Zieger, Cindy Greer, Stephanie Hatch, Joe Segal, Samuel Ji Blackford, Yu Ri Im, Vivek Chowdary, Taylor Ismali, Davide Danovi, Patrick A Lewis, James A Irving, Sunil Sahdeo, David A Lomas, Daniel Ebner, Christian Mueller, S Tamir Rashid.
      Alpha-1 antitrypsin deficiency (A1ATD) is a life-threatening condition caused by inheritance of the SERPINA1 'Z' genetic variant (PiZ) driving AAT protein misfolding in hepatocytes. There remain no approved medicines for this disease. Here, we report the results of a small molecule screen performed in patient derived iPSC-hepatocytes that identified Leucine-rich repeat kinase-2 (LRRK2) as a potentially new therapeutic target. Of the commercially available LRRK2 inhibitors tested, we identified CZC-25146, a candidate with favorable pharmacokinetic properties, as being capable of reducing polymer load, increasing normal AAT secretion, and reducing inflammatory cytokines in both cells and PiZ mice. Mechanistically, this effect was achieved through induction of autophagy. Our findings support the use of CZC-25146 and LRRK2 inhibitors in hepatic proteinopathy research and their further investigation as novel therapeutic candidates for A1ATD.
    DOI:  https://doi.org/10.1097/HEP.0000000000000969
  15. J Extracell Vesicles. 2024 Jul;13(7): e12469
    HaloTag display enables quantitative single-particle characterisation and functionalisation of engineered extracellular vesicles.
    Roxana E Mitrut, Devin M Stranford, Beth N DiBiase, Jonathan M Chan, Matthew D Bailey, Minrui Luo, Clare S Harper, Thomas J Meade, Muzhou Wang, Joshua N Leonard.
      Extracellular vesicles (EVs) play key roles in diverse biological processes, transport biomolecules between cells and have been engineered for therapeutic applications. A useful EV bioengineering strategy is to express engineered proteins on the EV surface to confer targeting, bioactivity and other properties. Measuring how incorporation varies across a population of EVs is important for characterising such materials and understanding their function, yet it remains challenging to quantitatively characterise the absolute number of engineered proteins incorporated at single-EV resolution. To address these needs, we developed a HaloTag-based characterisation platform in which dyes or other synthetic species can be covalently and stoichiometrically attached to engineered proteins on the EV surface. To evaluate this system, we employed several orthogonal quantification methods, including flow cytometry and fluorescence microscopy, and found that HaloTag-mediated quantification is generally robust across EV analysis methods. We compared HaloTag-labelling to antibody-labelling of EVs using single vesicle flow cytometry, enabling us to measure the substantial degree to which antibody labelling can underestimate proteins present on an EV. Finally, we demonstrate the use of HaloTag to compare between protein designs for EV bioengineering. Overall, the HaloTag system is a useful EV characterisation tool which complements and expands existing methods.
    Keywords:  HaloTag; extracellular vesicle; quantification; single vesicle
    DOI:  https://doi.org/10.1002/jev2.12469
  16. Amyotroph Lateral Scler Frontotemporal Degener. 2024 Jul 04. 1-4
    A novel TBK1 loss-of-function variant associated with ALS and parkinsonism phenotypes.
    Hiroya Naruse, Chifumi Iseki, Jun Mitsui, Jun Miki, Hikaru Nagasawa, Katsuro Kurokawa, Ryota Kobayashi, Hiroyasu Sato, Jun Goto, Wataru Satake, Hiroyuki Ishiura, Shoji Tsuji, Yasuyuki Ohta, Tatsushi Toda.
      Loss-of-function (LoF) variants in the TANK binding kinase 1 (TBK1) gene are implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. In this study, we present the first familial cases of ALS and parkinsonism associated with a novel TBK1 variant. We describe two siblings: one diagnosed with classical ALS and the other with a unique syndrome overlapping ALS and parkinsonism. Comprehensive clinical and imaging evaluations supported these diagnoses. Genetic analysis through whole-genome sequencing revealed a previously unknown heterozygous splice site variant in TBK1. Functional assessments demonstrated that this splice site variant leads to abnormal splicing and subsequent degradation of the mutated TBK1 allele by nonsense-mediated decay, confirming its pathogenic impact. Our findings suggest a broader involvement of TBK1 in neurodegenerative diseases and underscore the need for further research into TBK1's role, advocating for screening for TBK1 variants in similar familial cases.
    Keywords:  Amyotrophic lateral sclerosis; Parkinsonism; TBK1; loss-of-function variant
    DOI:  https://doi.org/10.1080/21678421.2024.2374374
  17. Biopreserv Biobank. 2024 Jul 03.
    Living Biobanks of Organoids: Valuable Resource for Translational Research.
    Wenqing Huang, Zhaoting Xu, Shuang Li, Junmei Zhou, Bing Zhao.
      The emergence of organoids is considered a revolutionary model, changing the landscape of traditional translational research. These three-dimensional miniatures of human organs or tissues, cultivated from stem cells or biospecimens obtained from patients, faithfully replicate the structural and functional characteristics of specific target organs or tissues. In this extensive review, we explore the profound impact of organoids and assess the current state of living organoid biobanks, which are essential repositories for cryopreserving organoids derived from a variety of diseases. These resources hold significant value for translational research. We delve into the diverse origins of organoids, the underlying technologies, and their roles in recapitulating human development, disease modeling, as well as their potential applications in the pharmaceutical field. With a particular emphasis on biobanking organoids for prospective applications, we discuss how these advancements expedite the transition from bench to bedside translational research, thereby fostering personalized medicine and enriching our comprehension of human health.
    Keywords:  living biobank; organoid; personalized medicine; translational research
    DOI:  https://doi.org/10.1089/bio.2023.0142
This page is being maintained by Thomas Krichel. It was last updated on 2024‒11‒25 at 17:34.