Biology subjects

Lin, L.-H.

Publications and source records attributed to Lin, L.-H..

5 recordsLinked to original sources

bioRxiv2026

Late radiation-induced brain injury is associated with functional cerebrovascular impairment without structural vessel loss in mice

Background: Radiation-induced brain injury (RIBI) is a major and often irreversible complication of cranial radiotherapy, affecting up to 90% of long-term survivors. Although cerebrovascular damage is a leading mechanistic hypothesis, whether chronic resting hypoperfusion occurs independently of structural vessel loss remains unknown. The aim of this study was to comprehensively characterize cerebrovascular, structural, and behavioral outcomes in a murine model of late RIBI to determine the primary drivers of vascular dysfunction. Methods: Adult C57BL/6 mice underwent 12 Gy whole-brain irradiation and were assessed one year later using laser speckle contrast imaging (LSCI), 7T MRI, immunohistochemistry, and behavioral testing. Results: Irradiated mice exhibited reduced pial flow velocity and flow area by LSCI, indicative of upstream cerebrovascular dysfunction, while bulk cortical tissue perfusion was preserved. This pial hypoperfusion was accompanied by cerebral atrophy, decreased T2* relaxation time and tissue iron deposition consistent with chronic microhemorrhage across multiple brain regions by MRI, blood-brain barrier disruption reflected by reduced ZO-1 and VE-cadherin expression, and increased anxiety-like behavior. Strikingly, capillary density, pericyte coverage, astrocyte endfeet, and NeuN-positive neuronal nuclei were all preserved. Conclusion: These findings demonstrate that functional vascular impairment - not structural vessel or neuronal loss - drives hypoperfusion and blood-brain barrier disruption in late RIBI. This dissociation between vascular dysfunction and rarefaction reframes the pathophysiology of RIBI and identifies endothelial dysfunction as a tractable therapeutic target for intervention.

neuroscience↗

bioRxiv2025

ASSET: A Framework for Decoding Aptamer Specificity of an Enriched Library by Next-Generation Sequencing of Experimental Samples

The SELEX process to identify RNA and DNA aptamers relies on sequencing selection rounds to detect highly specific aptamers through patterns of aptamer accumulation or enrichment. However, this approach infers rather than quantify aptamer specificity. Here we present a novel strategy for directly quantifying aptamer specificity within enriched libraries termed Aptamer Specificity Sequencing for Efficient Targeting (ASSET). The ASSET framework takes experimental samples and replicates testing the specificity of an aptamer library and prepares them for next-generation sequencing (NGS) with a known internal reference sequence. This enables robust data normalization, calculation of aptamer specificity scores with statistical significance, and the creation of specificity profiles of individual aptamers across multiple targets and non-targets. By integrating ASSET specificity scores with conventional selection round sequencing data, aptamers can be easily classified as true or false positives and negatives, allowing for easy separation of true positive aptamers. Compared to conventional methods for identifying aptamer candidates, such as measuring abundance or enrichment, ASSET specificity scores show a strong correlation with experimentally measured specificity. This supports ASSET as a more effective metric for selecting lead candidates following SELEX. ASSET is an easily implemented framework that accelerates the identification of highly specific aptamers, thereby expediting aptamer discovery for therapeutic and diagnostic applications.

molecular biology↗

bioRxiv2024

Identification of In Vivo Internalizing Cardiac-Specific RNA Aptamers

BackgroundThe pursuit of selective therapeutic delivery to target tissue types represents a key goal in the treatment of a range of adverse health issues, including diseases afflicting the heart. The development of new cardiac-specific ligands is a crucial step towards effectively targeting therapeutics to the heart. MethodsUtilizing an ex vivo and in vivo SELEX approaches, we enriched a library of 2-fluoro modified aptamers for ventricular cardiomyocyte specificity. Lead candidates were identified from this library, and their binding and internalization into cardiomyocytes was evaluated in both ex vivo and in vivo mouse studies. ResultsThe ex vivo and in vivo SELEX processes generated an aptamer library with significant cardiac specificity over non-cardiac tissues such as liver and skeletal muscle. Our lead candidate aptamer from this library, CA1, demonstrates selective in vivo targeting and delivery of a fluorophore cargo to ventricular cardiomyocytes within the murine heart, while minimizing off-target localization to non-cardiac tissues, including the liver. By employing a novel RNase-based assay to evaluate aptamer interactions with cardiomyocytes, we discovered that CA1 predominantly internalizes into ventricular cardiomyocytes; conversely, another candidate CA41 primarily binds to the cardiomyocyte cell surface. ConclusionsThese findings suggest that CA1 and CA41 have the potential to be promising candidates for targeted drug delivery and imaging applications in cardiac diseases.

bioengineering↗

bioRxiv2024

MIRO1 controls energy production and proliferation of smooth muscle cells

BackgroundThe outer mitochondrial Rho GTPase 1, MIRO1, mediates mitochondrial motility within cells, but implications for vascular smooth muscle cell (VSMC) physiology and its roles in vascular diseases, such as neointima formation following vascular injury are widely unknown. MethodsCarotid ligation was performed in an in vivo model of selective Miro1 deletion in smooth muscle cells. VSMC proliferation during the cell cycle and molecular mechanisms of smooth muscle cell proliferation were explored in cultured aortic VSMCs by imaging mitochondrial positioning and cristae structure and assessing the effects on ATP production, metabolic function, and interactions with components of the electron transport chain (ETC). MIRO1 expression was also analyzed in human coronary arteries, and its function was assessed via knockdown in human coronary artery VSMCs. ResultsMIRO1 was highly expressed in VSMCs within human atherosclerotic plaques. MIRO1 facilitated VSMC proliferation and neointima formation by regulating mitochondrial positioning and PDGF-stimulated ATP production and respiration, critical for cell-cycle progression at G1/S. Deletion of Miro1 disrupted mitochondrial cristae structure, diminished ETC complex I activity, and impaired supercomplex formation. Notably, restoring MIRO1 function with expression of wild type MIRO1 recovered proliferation and ATP production and respiration, whereas a mutant lacking EF hands, which are essential for mitochondrial motility, only partially rescued these effects. MIRO1 knockdown in human coronary artery VSMCs confirmed its pivotal role in mitochondrial function and VSMC proliferation. ConclusionsThis study highlights two key mechanisms by which MIRO1 regulates VSMC proliferation. First, it maintains ATP synthesis by preserving mitochondrial cristae integrity. Second, its Ca2+-dependent EF hands enable ATP-dependent mitochondrial positioning. By linking mitochondrial motility and energy production to VSMC physiology, these findings position MIRO1 as a critical regulator of vascular remodeling and a potential target for therapeutic interventions.

cell biology↗

bioRxiv2022

Gut Microbiome Contributes to Cold-climate Adaptation in Lizards

The metabolic cold-climate adaption hypothesis predicts that animals from cold environments have relatively high metabolic rates compared with their warm-climate counterparts. However, studies testing this hypothesis are sparse. Here, we compared gut microbes between two cold-climate lizard species of the genus Phrynocephalus to test the hypothesis that gut microbiota can help lizards adapt to cold environments by promoting metabolism and absorption efficiency. We kept lizards at 24{degrees}C and 30{degrees}C for 25 d, and then collected their fecal samples to analyze and compare the microbiota based on 16S rRNA gene sequencing technology. The gut microbiota was mainly composed of Proteobacteria, Firmicutes, Bacteroidetes, and Verrucomicrobia at the phylum level in both species (Proteobacteria > Firmicutes > Verrucomicrobiota in P. erythrurus, and Bacteroidota > Proteobacteria > Firmicutes in P. przewalskii). Further analysis revealed that the gut microbiota contributed to the hosts cold adaptation, but with differences in the relative abundance of these contributory bacteria between the two species. KEGG analysis revealed that the gut microbiota primarily played roles in metabolism, genetic information processing, cellular processes, and environmental information processing in both species. Furthermore, genes related to metabolism were more abundant in P. erythrurus at 24 {degrees}C than in other species x temperature combinations, indicating the role of gut microbiota in long-term cold-climate adaptation. Our finding that gut microbiome contributes to cold-climate adaptation in both species but more evidently in P. erythrurus using colder habitats than P. przewalskii throughout a year confirms the gut microbiotas role in the cold-climate adaptation in lizards. IMPORTANCEThis study proves that temperature affects the composition and relative abundance of the gut microbiota in two Phrynocephalus lizards in a species-specific manner. Both species harbor specific gut microbiota with significant roles in cold-climate adaptation. Specifically, P. erythrurus has a higher Proteobacteria ratio and relative abundance of metabolism-related microbial genes in the gut than P. przewalskii. Given that P. erythrurus uses colder habitats than P. przewalskii throughout a year, these results suggest that gut microbiota contributes to cold-climate adaptation in both species lizards but more evidently in P. erythrurus. This study provides evidence linking gut microbiome with cold adaptation. The co-evolution mechanism between gut microbiota and their hosts in extreme environments will provide new insights into animal adaptation.

ecology↗