Search bioRxiv⌕ Search

Biology subjects

Lin, J.

Publications and source records attributed to Lin, J..

At least 55 records · Page 3Linked to original sources

The Euler Characteristic Transform Enables Classification of Complex Plant Shapes and Prediction of Leaf Venation from Blade Geometry

(1) RationaleQuantifying and predicting plant morphology is central to understanding development and evolution, yet many plant forms lack homologous features required for traditional morphometrics. We apply the Euler Characteristic Transform (ECT), an injective descriptor from topological data analysis, to encode 2D plant shapes. The ECT converts contours into image-like representations that preserve shape information while enabling deep learning. (2) MethodsWe computed ECTs for large datasets of leaf and pavement cell shapes and used convolutional neural networks (CNNs) for classification. We also trained CNNs to approximate the inverse mapping, predicting leaf shape masks from radial ECTs. (3) Key resultsECT-based models achieved high classification accuracy, surpassing previous approaches on millions of herbarium-derived leaves. Notably, grapevine leaf venation was predicted from blade geometry alone, demonstrating that vascular structure is encoded in the outline. (4) Main conclusionThe ECT provides a compact, information-preserving representation of biological shape that integrates naturally with deep learning. It enables both accurate classification and predictive reconstruction, revealing latent morphological information and offering new opportunities to study plant form across scales.

plant biology↗

SIK3-HDAC4-Warts Axis Functions as a Gatekeeper of Neural Stem Cell Reactivation in Drosophila

A delicate balance between the quiescent and proliferative states of neural stem cells (NSCs) is important for neurogenesis and homeostasis. Histone deacetylase 4 (HDAC4) variants are associated with neurodevelopmental disorders, however, its role in early brain development remains elusive. In this study, we demonstrate that Drosophila HDAC4 plays a crucial role in neural stem cells (NSCs) reactivation and brain development. Depletion of HDAC4 results in notable defects in NSC reactivation, while its overexpression leads to premature reactivation. HDAC4 is SUMOylated at Lys902, which enhances its protein stability by preventing HDAC4 from undergoing ubiquitin-proteasome-mediated degradation. Moreover, phosphorylation of HDAC4 by salt-inducible kinase 3 (SIK3), an AMPK-related kinase, allows cytoplasmic localization of HDAC4 and enhances the association between HDAC4 and Warts, a core kinase of the Hippo pathway. This HDAC4-Wts association inhibits Warts activity, and in turn, the inactivation of the Hippo pathway, triggering NSC reactivation. Finally, genetic epistasis experiments support the SIK3-HDAC4-Warts axis during NSC reactivation. In conclusion, our findings identify HDAC4 as a molecular switch that integrates SUMOylation, ubiquitination, and the Hippo pathway to govern NSC reactivation.

neuroscience↗

Modulation of Oncogenic KRAS Signaling by Branched Actin-driven Cell Membrane Protrusions

For over three decades, we have known that oncogenic RAS alters the actin cytoskeleton organization and cell surface morphology1,2. RAS activates the GTPase RAC1, which triggers the growth of branched actin networks to promote cell membrane protrusions3,4. In melanoma, the hyperactive RAC1 mutant, Rac1P29S, was recently shown to drive extended lamellipodia, which then empower cell proliferation through sequestration and localized inhibition of the merlin tumor suppressor5. This discovery illustrates cell morphological programs not only as outputs but also as regulators of human oncogenic signals. Hence, we wondered whether the pronounced branched actin-driven membrane protrusions (BAMPs) downstream of oncogenic RAS are not mere outputs of RAS signaling but rather an active component in mediating the oncogenic penetrance of RAS mutants. We used volumetric light sheet microscopy and biochemical approaches to investigate the role of BAMPs in regulating the molecular signaling of oncogenic KRAS in pancreatic and lung cancer models. We found that elevated BAMP formation regulated the interaction of oncogenic KRAS with downstream effectors, specifically with the RAC1 GEF TIAM1. This implies that BAMPs amplify their own upstream regulators in a positive feedback. This meritorious cycle upregulates cyclin D1 expression by inactivating the merlin tumor suppressor, independently of the mitogen activated protein kinase pathway (MAPK). In the absence of BAMPs, cells carrying oncogenic KRAS mutations are unable to attain their full penetrance in proliferation. Overall, this work unveils the long-overlooked role of branched actin-driven cell morphology in the functionalization of KRAS mutants as potent oncogenes.

cancer biology↗

Engineering age-adaptive mRNA lipid nanoparticle cancer vaccines via reprogramming systemic gene expression

Lipid nanoparticle (LNP)-based mRNA vaccines have transformed cancer immunotherapy, yet their efficacy in older individuals, who represent the majority of cancer patients, remains poorly understood. Here, we uncover a critical and previously underappreciated barrier to mRNA LNP cancer vaccine performance in aged hosts: impaired systemic transgene expression. Using the SM-102 mRNA LNPs as a benchmark formulation, we show that while local immune activation and antigen presentation at the injection site and draining lymph nodes remain largely intact with age, transgene expression in peripheral organs, including the liver, lungs, and spleen, is markedly reduced. This deficit limits the magnitude and durability of CD8 and CD4 T cell responses and substantially compromises tumour control. Transcriptomic profiling further reveals that attenuated transgene expression parallels broad attenuation of antigen processing, presentation, and activation pathways in immune cells from aged animals, implicating impaired systemic mRNA translation as a central driver of the downstream immune defects and antitumour efficacy loss. Building on these insights, we identify a rationally selected LNP formulation that reestablishes distal antigen expression across age groups as an engineering strategy to revive T cell immunity, achieving full rescue of therapeutic efficacy in aged mice without additional intervention. Together, these findings establish systemic mRNA translation as a tuneable lever of vaccine performance and highlight that optimizing LNP formulations to sustain systemic transgene expression across age groups may enable next-generation, age-adaptive mRNA vaccines for cancer and other diseases of aging.

bioengineering↗

Physical theory of epigenetic memory and its biological implications

Epigenetic memory encoded via histone modifications must remain robust across cell divisions. However, the physical rules governing how chromatin architecture regulates the maintenance and reorganization of epigenetic patterns remain unclear. Here, we develop a spreading-writing-erasing theory of epigenetic memory incorporating chromatin compartmentalization, enzyme limitation, and mark spreading. Our model demonstrates that self-sustaining epigenetic patterns emerge naturally without sequence insulators, while the establishment and erasure of heterochromatic compartments is governed by finite thresholds. Crucially, we uncover that the power-law exponent of chromatin contact probability dictates memory stability by buffering against spontaneous compartment collapse or expansion. Our theory also predicts progressive compartment fusion over cell generations, setting an upper limit of about 60 generations before senescence for human cells and explaining Hi-C map dynamics in senescent cells. Our theory also predicts a positive correlation between the contact-decay exponent and organismal complexity across species, supported by analysis of Hi-C maps across 873 species.

cell biology↗

Generation of Genetically Identical Mammalian Oocytes from Parthenogenetic Double-Haploid Embryonic Stem Cells

Generating genetically identical mammalian oocytes is challenging due to stochastic meiotic recombination. Here, we established parthenogenetic double-haploid embryonic stem cells (PG-DhESCs) possessing complete homozygosity. By employing blastocyst complementation in Prdm14-deficient embryos, we generated chimeric females that produced oocytes derived exclusively from these donor cells. Fertilization of these oocytes yielded viable, fertile, maternally semi-cloned (MSC) mice of both sexes. Although DNA methylation was largely restored during gametogenesis, subtle epigenetic defects correlated with increased body weight in MSC offspring. This study establishes a robust platform combining PG-DhESCs with blastocyst complementation to generate isogenic mammalian oocytes, overcoming traditional limitations in mammalian cloning.

developmental biology↗

CGRP receptor-expressing neurons in the central amygdala contributes to injury-induced pain hypersensitivity

The central nucleus of amygdala (CeA) comprises diverse populations of neurons, forming a complex network responsible for regulating various behavioral responses. Among these, neurons expressing calcitonin gene-related peptide receptors (CGRPR) have emerged as key players in CGRP neuropeptide-mediated pain modulation. While previous studies emphasize CGRPs key role in synaptic plasticity and its connection with pain behavior in the CeA, the precise functional attributes and contributions of CeA-CGRPR-expressing neurons in pain processing remain elusive. This study reveals the co-localization of CGRPR-expressing neurons in the CeA with phosphorylated extracellular signal-regulated kinase (pERK), a marker indicating pain plasticity, in a neuropathic pain model. Electrophysiological assessments of these neurons in slice preparations unveiled heightened intrinsic excitability after sciatic nerve cuff implantation, contingent upon their rostro-caudal positioning within the CeA. Furthermore, our behavioral experiments using chemogenetic inhibition of CeA-CGRPR neurons demonstrated the ability to reverse nerve injury-induced hypersensitivity. Conversely, activating these neurons induced pain-related hypersensitivity even in the absence of injury. Our findings also highlight a sex-specific role of CeA-CGRPR neurons in formalin-induced spontaneous pain response. Collectively, these data reinforce the involvement of CeA-CGRPR neurons in pain processing, contributing to a better understanding of how neural circuits are affected in persistent pain conditions. Significance StatementOur study shows the role of CGRPR-expressing neurons within the CeA during pain processing. Using a cuff-implanted neuropathic mouse model, we discovered that CGRPR-expressing neurons co-localize with phosphorylated extracellular signal-regulated kinase (pERK), a hallmark of pain plasticity, in both male and female mice. Furthermore, our electrophysiological investigations reveal that posterior CeA-CGRPR neurons exhibit increased excitability following sciatic nerve cuff implantation. Importantly, we demonstrate that CeA-CGRPR neurons exert bidirectional effects on pain behavior in mice, irrespective of sex differences in nerve injury-induced pain responses while showing sex-specific spontaneous pain responses in the formalin-induced model. These findings show the role of CeA-CGRPR neurons in pain modulation, underscoring their potential significance in understanding and addressing persistent pain conditions.

neuroscience↗

Adaptive coupling of chromosomal inversions with multilocus ecological, mating-bias, and hybrid-incompatibility genotypes facilitates sympatric speciation

Chromosomal inversions (CIs) are widespread structural variants that suppress recombination and maintain favorable allele combinations as tightly linked supergenes. They are increasingly recognized for their contributions to local adaptation and speciation, yet their role in sympatric speciation under ongoing gene flow remains unresolved. Here, we develop mathematical models and computer simulations to examine how polygenic architectures and CI invasion jointly shape the evolution of reproductive isolation (RI) in a two-niche model under disruptive ecological selection. Extending previous frameworks, we incorporate multilocus ecological traits, multilocus mating-bias traits, and multilocus hybrid-incompatibility genotypes, and evaluate the invasion fitness and evolutionary consequences of CIs capturing different combinations of locally adaptive alleles. We show that the evolutionary impact of CI invasion depends critically on how polygenic trait structure influences hybrid production and the strength of barrier mechanisms. Increasing the number of ecological or hybrid-incompatibility loci strengthens disruptive ecological selection and postzygotic incompatibility selection by generating more unfit hybrids, whereas increasing the number of mating-bias loci weakens premating selection because mating-bias hybrids remain viable within the same niche. Accordingly, CIs capturing ecological or hybrid-incompatibility alleles tend to reduce the effective number of loci, diminish hybrid loss, and weaken existing RI, whereas CIs capturing mating-bias alleles strengthen premating isolation by reducing the effective number of mating-bias loci. Importantly, CIs that couple alleles across distinct barrier mechanisms exhibit elevated invasion fitness and generate synergistic reinforcement and positive feedback among premating isolation, postmating isolation, and ecological divergence. These findings reconcile contrasting theoretical predictions by demonstrating that CIs can either facilitate or constrain sympatric speciation depending on how they reshape effective locus number and barrier coupling, and provide a unified framework for understanding how structural genomic variants interact with polygenic architectures to influence the origin and stability of reproductive isolation.

evolutionary biology↗

A lateral linker histone binding mode scaffolds dinucleosome stacking in chromatin fibers

Linker histones are essential for chromatin compaction, yet how they contribute to higher-order fiber assembly remains poorly understood. Here, we determined cryo-electron microscopy structures of Arabidopsis dodeca-nucleosome fibers containing distinct H2A/H3 variants and linker histone H1.3, revealing a noncanonical binding mode that a laterally positioned H1.3 connects the acidic patch of one nucleosome and the DNA of the neighboring nucleosome, thereby scaffolding dinucleosomes into two-start chromatin fibers. This noncanonical binding mode is structurally conserved when H1.3 is replaced by Gallus gallus H5. Furthermore, incorporation of H2A.W and H3.3 further induces back-to-back fiber dimerization. Cryo-electron tomography and in vivo cross-linking mass spectrometry analyses support the physiological relevance of H1 lateral engagement. Our findings establish that linker histones act as active architectural scaffolds in higher-order chromatin organization.

molecular biology↗

Subdiffusive random growth of bacteria

While the regulation of bacterial cell size is widely studied across generations, the stochastic nature of cell volume growth remains elusive within a cell cycle. Here, we investigate the fluctuations of cell volume growth and report a subdiffusive random growth. Specifically, the mean square displacement of the logarithmic volume scales as {Delta}t with an anomalous exponent {approx} 0.27. This low exponent implies strong negative temporal correlations in growth rate noise on timescales of minutes, which are significantly faster than those of gene expression dynamics. We attribute this phenomenon to the physical mechanics of the cell wall. By modeling the cell wall as a complex viscoelastic material with power-law-distributed relaxation times, we successfully recapitulate the observed subdiffusive behavior. Our results suggest that it is the heterogeneous cell-wall viscoelasticity, rather than biological regulatory programs, that governs the short-timescale fluctuations of bacterial growth.

biophysics↗

Efficient Reprogramming of the Epiblast Enables the Generation of Cloned Mice

Somatic cell nuclear transfer (SCNT) can generate viable mammals despite pervasive epigenetic abnormalities in cloned embryos, yet the mechanism underlying this paradox remains unclear. Here we show, using single-cell transcriptomics, that efficient reprogramming occurs exclusively in the epiblast (EPI), but the not primitive endoderm (PrE) of late mouse SCNT blastocysts. Integrating this with previous findings of trophectoderm (TE) aberration, we propose the EPI as the sole effectively reprogrammed lineage. By innovatively employing a Lego-like multi-lineage embryonic aggregation approach, where the extra-embryonic lineages were replaced with fertilization-derived counterparts, we demonstrated that SCNT-derived EPI inherently possesses full-term developmental potential nearly identical to that of fertilized EPI (20.5% vs. 21.8% birth rate), functionally confirming its effective reprogramming. Our study uncovers a lineage-specific asymmetric reprogramming mode where the EPI specifically achieves effective reprogramming, thereby constituting the deterministic basis for cloned animal generation. This work also provides a versatile strategy for investigating lineage potency and function.

developmental biology↗

Co-evolution of Oncogenic KRAS Signaling and LILRBhigh Macrophages Drives Pancreatic Cancer Recurrence

Pancreatic ductal adenocarcinoma (PDAC) frequently recurs after surgical resection, indicating that residual disease is sustained by coordinated tumor-microenvironment interactions. To define the biological basis of recurrence, we leverage large-scale clinical data from 2,710 patients, deeply characterized multi-omics profiling (whole-exome, bulk RNA, and single-nucleus sequencing) of 36 matched primary and locally recurrent PDACs, an in-house multiplex spatial imaging cohort of 190 patients, and extensive public datasets. Recurrent tumors were characterized by increased KRAS mutant allele dosage and reinforced KRAS signaling, accompanied by expansion of basal-like malignant cell states. In parallel, we identified an immunosuppressive macrophage population marked by high LILRB expression that spatially co-localized with KRAS-activated tumor cells. Functional studies showed that LILRB4+ macrophages enhanced tumor cell plasticity and progression, whereas inhibition of macrophage LILRB4 suppressed these phenotypes. Notably, a first-in-class human anti-LILRB4 antibody reduced macrophage-driven tumor traits, and dual targeting of KRAS signaling and LILRB4 achieved superior tumor control in macrophage-containing mouse models. These findings reveal a co-evolved tumor-immune niche underlying PDAC recurrence and nominate the KRAS-LILRB4 axis as a therapeutic vulnerability.

cancer biology↗

Genome-wide maps of transcription factor footprints identify noncoding variants rewiring gene regulatory networks with varTFBridge

Genome-wide association studies have identified millions of noncoding loci linked to human traits, yet how these variants alter gene regulation remains a major challenge, particularly for rare variants where whole-genome sequencing cohorts and high-resolution functional annotations remain limited. Here we show that single-molecule deaminase footprinting (FOODIE) in K562 cells captures up to 103-fold heritability enrichment for erythroid traits despite covering 0.12% of the genome. We introduce varTFBridge, integrating FOODIE footprinting with AlphaGenome variant effect prediction to identify causal noncoding variants altering transcription factor (TF)-mediated regulation. Applied to 490,640 UK Biobank genomes across 13 erythrocyte traits, varTFBridge prioritises 113 high-confidence regulatory variants (104 common, 9 rare), encompassing 2,173 linkages along the variant-TF binding- gene-trait cascade across 64 TFs and 108 genes. varTFBridge recapitulates rs112233623 and resolves its mechanism: GATA1/TAL1 co-binding disruption at a CCND3 enhancer altering red blood cell count and volume.

bioinformatics↗

Mitochondria Organize Gap Junctions to Enable Neural Circuit Scaling

Neural circuits must maintain function during dramatic growth-driven expansion. While chemical synapses can be independently reorganized in pre- and postsynaptic neurons, gap junctions physically couple partner cells, requiring coordinated spatial redistribution--a process whose mechanisms remain unknown. Using the C. elegans mechanosensory circuit, we show that gap junctions undergo stereotyped developmental reorganization, transitioning from clustered to dispersed configurations as partner neurites elongate. Forward genetic screening identified RIC-7, a mitochondrial adaptor protein, as essential for this redistribution. Loss of RIC-7 prevents gap junction dispersal, trapping partner neurites at clustered sites and severely impairing electrical transmission. Mechanistically, RIC-7 directs mitochondrial positioning to gap junctions, where mitochondria recruit the microtubule-organizing component PTRN-1 to locally enhance microtubule dynamics. Disrupting mitochondrial transport (miro-1;mtx-2 mutants) or PTRN-1 function phenocopies ric-7 defects, while metabolic dysfunction does not, establishing a non-metabolic role for mitochondria in organizing synaptic architecture. Remarkably, RIC-7 expression in one neuron alone suffices to restore gap junction and partner neurite distribution across both coupled cells. Structural modeling suggests mechanical adhesion between docked hemichannels enables this coordinated reorganization. These findings establish mitochondria as mobile organizers of synaptic spatial architecture, demonstrate that proper junction distribution is essential for electrical transmission efficacy, and reveal how cell-autonomous mechanisms can achieve bilateral structural coordination during circuit growth.

neuroscience↗

A human gut-BBB-brain microphysiological model for studying neurodegenerative diseases

The gut-brain axis has emerged as a crucial factor in neurodegeneration, with growing evidence linking gut dysbiosis and metabolic dysfunction to Alzheimers disease (AD) progression. Unfortunately, the lack of human-relevant in vitro models limits our ability to effectively explore the mechanism of this axis. To address this gap, we have developed a human induced pluripotent stem cell (iPSC)-derived gut-blood-brain barrier (BBB)-brain microphysiological system that enables systematic investigation of gut-brain interaction in the context of AD under controlled conditions. Our findings reveal that the interaction between gut and brain organoids can promote the maturation of brain organoids, making them more similar to their physiological characteristics in vivo. Additionally, co-culture gut and brain organoids better recapitulates the pathological features of AD. We also discovered that gut organoids of AD can trigger neurodegenerative disease manifestations in healthy brain organoids. In summary, our microphysiological system provides a novel and versatile in vitro platform for studying the interaction between the gut and brain in neurodegenerative diseases.

neuroscience↗

Efficient derivation and transcriptional characterization of mouse extra-embryonic endoderm stem cell lines generated by somatic cell nuclear transfer

Somatic cell nuclear transfer (SCNT) holds great promise for regenerative medicine and agriculture, but its application is severely hampered by low efficiency, primarily attributable to aberrant epigenetic reprogramming. Although embryonic stem cells (ESCs) and trophoblast stem cells (TSCs) have been successfully derived from cloned embryos, an in vitro counterpart of the primitive endoderm (PrE) lineage has remained unavailable. To address this gap, this study reports the first successful establishment of extra-embryonic endoderm stem cell lines (XENs) from mouse SCNT-derived blastocysts (NT-XENs). Under conventional culture conditions, NT-XENs were generated from hybrid B6D2F1 blastocysts at a high efficiency of 55%, comparable to that of fertilization-derived XEN lines (FD-XENs, 50%), whereas derivation from inbred C57BL/6J SCNT-derived blastocysts was markedly lower (12.5%). Immunofluorescence and NanoString multiplex gene expression profiling confirmed that NT-XENs robustly expressed specific marker genes for PrE/XENs (e.g., Gata4, Gata6, Sox17), while exhibiting negligible or absent expression of pluripotency and trophoblast markers. Based on NanoString assay data, NT-XENs and FD-XENs shared highly similar global gene expression patterns, yet also exhibited some nonnegligible differences, exemplified by the differentially expressed genes (DEGs) Pecam1, Gtl2, Thbd and Xlr3b, which may suggest that the NT-XENs resided in a more differentiated state (potentially biased toward parietal endoderm (PE)) and retained SCNT-specific epigenetic imprinting errors, including aberrant X-chromosome inactivation and dysregulation of imprinted domains. In summary, this study successfully establishes NT-XEN cell lines, providing a valuable in vitro model for investigating the reprogramming scenarios of PrE lineage in SCNT and offering novel insights into the mechanisms underlying developmental failure of cloned embryos.

developmental biology↗

Donor-specific assemblies enhance somatic structural variant detection in complex genomic regions

Structural variants (SVs) contribute substantially to genomic variation and disease, but detecting somatic SVs (sSVs) remains difficult due to reference bias, mosaicism, and enrichment in repetitive regions. Linear reference genomes, like GRCh38 and CHM13, do not fully capture individual genomic structure, which can obscure true somatic variation. Donor-specific assemblies (DSAs) generated from the same genome where sSVs are being assayed provide a personalized alternative, yet their performance for sSV detection has not been systematically assessed. As part of the Somatic Mosaicism across Human Tissues (SMaHT) Network, we benchmark a DSA for sSV discovery in the COLO829 melanoma cell line with a matched normal sample from the same individual. We compare sSV detection across GRCh38, CHM13, and the COLO829BL_DSA using three different sSV callers (Delly, Severus, and Sniffles2) and sequence data from multiple long-read platforms. The COLO829BL_DSA identifies 1.8-fold more manually validated sSVs than linear references, in regions both shared with GRCh38 and CHM13 and unique to the COLO829BL_DSA. Variants detected only with the COLO829BL_DSA are often found in satellite and other repeat-rich regions that are difficult to resolve using standard references. In addition, several COLO829BL_DSA-specific sSVs are located in genes, some of which are associated with cancer. Overall, these results underscore the utility of DSAs in improving sSV detection.

genomics↗

Heterogeneous and specific localization of mRNAs at dendritic spine and its dependence on microtubule entry and kinesin motor

Local mRNA translation near neuronal synapses is crucial for structural plasticity during brain development and memory formation. Whereas mRNAs are transported from soma to dendrites as individual ribonucleoprotein (RNP) granules, how do they subsequently localize at synapses is unclear. In particular, whether different dendritic mRNAs might display differential synaptic localization and interact with specific postsynaptic proteins remains unexplored. Here we found that two well-known dendritic mRNAs, Actb and Camk2a, are targeted to different subdomains of dendritic spines and delivered to different spine populations after synaptic stimulation. Surprisingly, the induced mRNA localization to dendritic spines is associated with microtubule (MT) entry and specifically requires the MT-associated motor KIF5A. Proximity-labelling proteomics further identifies distinct postsynaptic proteins enriched by Actb and Camk2a, from which we uncover the cytoskeletal protein Septin7 to be co-localized with the two mRNAs at different subdomains of dendritic spines. Our study reveals an unexpectedly specific mRNA localization mechanism at synapses and demonstrates proximity-labelling as a useful approach to map their synaptic partners.

neuroscience↗