Biology subjects

Liang, P.

Publications and source records attributed to Liang, P..

At least 19 recordsLinked to original sources

bioRxiv2026

Ultra-high-efficacy Wireless Neuromodulation Using a Conductive Ring Interface

An ab-initio physics study, supported with a COMSOL-based numerical simulation, presents a concept to enable wearable, highly targeted, and ultra-high-efficacy neuromodulation. A target site is defined by a relatively small gap of a miniaturized ring-shaped conductive interface implant. If the implant conductivity significantly exceeds the conductivity of the biological tissue, a highly localized electromotive force (emf) is induced in the ring upon application of an alternating current (AC) magnetic field. This emf generates an electric field in the gap, inducing local neural stimulation of the targeted site, e.g., translated to a nerve segment using circumferential cuffs. The wirelessly controlled autonomous stimulator implant acts as a low-resistance nearly-closed path to generate a highly localized electric field on the order of 50 V/m in the gap via application of a 1-kA/m AC magnetic field at a frequency of 10 kHz. The magnetic field is at least three orders of magnitude smaller than the magnetic field required to generate the same electric field strength via conventional transcranial magnetic stimulation. Therefore, this high-resolution, orientation-specific ring neuromodulation system could enable a highly localized, energy-efficient wearable device with significant preclinical and clinical potential.

bioengineering↗

bioRxiv2026

A cross-kingdom interactome predicted by AlphaFold3 reveals a DNF2-centered interface required for symbiotic accommodation

Legumes convert atmospheric nitrogen into ammonium through symbiotic bacteria housed in root nodules, yet the molecular interactions between rhizobial and host proteins inside nodules remain poorly understood. Here we employed AlphaFold3 to construct a cross-kingdom interactome between Medicago truncatula and its symbiont Sinorhizobium meliloti. Screening more than 217,000 protein pairs yielded 7,137 putative interactions, providing a valuable resource for the broader symbiosis community. Within this network, we focused on DEFECTIVE IN NITROGEN FIXATION 2 (DNF2), a host protein required for rhizobial persistence within nodules. We showed that DNF2 localizes to the peribacteroid space and associates with previously uncharacterized secreted rhizobial proteins (SRPs), suggesting it may function as a hub for host-symbiont communication. Notably, knockout of two DNF2-interacting proteins, SRP86 and SRP485, results in white, nitrogen-fixation-deficient nodules with abnormal symbiosomes and elevated expression of senescence-associated genes, closely phenocopying the dnf2 loss-of-function mutant. Together, our findings define a DNF2-SRP molecular framework underlying symbiotic accommodation, and illustrate the potential of AI-guided interactome mapping to uncover molecular mechanisms of plant-microbe interactions with relevance to sustainable agriculture.

plant biology↗

bioRxiv2026

Revisiting the diversity of secondary endosymbionts in the major pest oat aphid, Rhopalosiphum padi

There is disagreement on whether secondary endosymbionts are found in the major cereal pest aphid, Rhopalosiphum padi. Some papers report a diversity of secondary bacterial endosymbionts while others have failed to find evidence of these bacteria in this species. Here we revisit this issue by summarizing the relevant literature and through additional sampling of the species in Australia, China and Denmark using a combination of molecular approaches. We find a general absence of secondary endosymbionts beyond the obligate endosymbiont Hamiltonella defensa in R. padi. While the inconsistency in survey results may reflect rapid changes in endosymbiont turnover in populations and/or the impact of ecological factors such as host plant type on endosymbiont diversity, we are concerned that technical issues may be at least partly responsible for inconsistencies in the literature. This leads us to emphasize the importance of multiple sources of evidence required to establish and characterize endosymbiont infections, including PCR and qPCR assays, DNA Sanger sequencing and 16SrRNA gene metabarcoding. We note that several major aphid pests show a low incidence of secondary endosymbionts which raises issues about the importance of these endosymbionts in aphids that constitute pests, even though endosymbionts can in some cases increase host fitness and therefore pest impact.

microbiology↗

bioRxiv2026

Structures and molecular mechanisms of RAD54B in modulating homologous recombination

Genome stability is essential for cellular viability yet constantly threatened by endogenous and exogenous DNA-damaging agents. Among these, DNA double-strand breaks (DSBs) are particularly harmful and in S/G2 phases are faithfully repaired through homologous recombination (HR), a high-fidelity pathway utilising homologous sequences in sister chromatin. The RAD51 recombinase forms nucleoprotein filaments on single-stranded DNA (ssDNA) to mediate homology search, strand invasion and subsequent D-loop formation that leads to DNA synthesis and repair. The efficiency of HR depends on precise regulation of RAD51 filament dynamics by accessory factors, including RAD54 and RAD54B, which belong to the SWI2/SNF2-family DNA translocases. While RAD54 is well-characterized, RAD54Bs molecular functions remain poorly understood. Here, we define RAD54Bs role in HR using cryo-electron microscopy, mutagenesis, biochemical and cellular assays. We show that RAD54B stabilizes RAD51-DNA filaments, inhibits RAD51 ATPase activity, and promotes strand invasion, D-loop formation and strand exchange. The N-terminal domain (NTD) alone supports filament stabilization and strand exchange, while the C-terminal ATPase domain is required for D-loop formation. Structural and biochemical analyses reveal three RAD51-interacting sites within the NTD and a unique domain ({beta}-domain) that bridges RAD51 protomers and contacts donor dsDNA. This {beta}-domain also regulates RAD54Bs ATPase activity and higher-order oligomer organization on dsDNA. Cellular assays reveal that the NTD RAD51-interacting sites as well as the {beta}-domain are required for repairing camptothecin-induced DSBs by HR in human cells. Our findings uncover a modular architecture and mechanistic framework for RAD54B function in HR, highlighting its critical role in genome maintenance. HighlightsO_LIcryoEM structure of RAD54B in complex with RAD51-DNA complex C_LIO_LIRAD54B uses three sites to interact with RAD51, including a previously unrecognised {beta}-domain that bridges distal RAD51 protomers. C_LIO_LIThe {beta}-domain plays multiple crucial roles including regulating filament stability, RAD54B ATPase activity and RAD54B higher order assembly on DNA. C_LIO_LIRAD54B employs a modular mechanism, with the N-terminal region engaing and stabilising RAD51 filaments, capturing of the homologous strands, whereas the ATPase motor domainrequired for homology search and strand invasion. C_LIO_LIRAD54B N-terminus and {beta}-domain are essential for HR-mediated repair of camptothecin-induced breaks in human cells. C_LI

biochemistry↗

bioRxiv2026

Exploring Multi-Scale Local and Global Features in Whole Slide Images Using State Space Models

Whole slide image (WSI) classification is crucial in computational pathology, yet the gigapixel scale of WSIs makes it challenging to extract discriminative and compact WSI-level features for disease diagnosis. In this paper, we propose MambaWSI, a novel method that leverages the state space model (SSM) for WSI classification by exploring multi-scale local and global features. Unlike existing approaches that sequentially traverse WSI tiles and rely on vanilla SSMs for long-range dependency modeling, we exploit a traversal strategy in a higher-dimensional discrete space that preserves spatial proximity, enabling a first-local-then-global feature extraction process. Furthermore, to align with the clinical workflow of pathologists when examining WSIs at multiple scales, we propose a two-stage hierarchical fusion strategy: inter-scale feature alignment and aggregation, followed by attention-based fusion across magnifications, integrating complementary information from multiple magnifications. Experiments on two datasets demonstrate that MambaWSI outperforms state-of-the-art methods in classification performance.1

pathology↗

bioRxiv2026

Cannabidiol Inhibits PIEZO Channels to Mitigate Red Blood Disorders

Hyperactivity of the mechanosensitive ion channel PIEZO1 promotes pathologic Ca{superscript 2} overload in red blood cells (RBCs), driving dehydration, TMEM16F-dependent phosphatidylserine (PS) exposure, microparticle shedding, and increased thrombotic and vaso-occlusive risks in hereditary xerocytosis (HX) and sickle cell disease (SCD). However, clinically deployable PIEZO inhibitors to treat these blood disorders are lacking. Here we report that cannabidiol (CBD), a non-psychoactive cannabinoid commonly used in SCD patients for pain management, inhibits PIEZO1 activity and restores aberrant mechanotransduction in HX and SCD RBCs. Micromolar concentrations of CBD blocks PIEZO1 currents and suppresses PIEZO1-mediate Ca{superscript 2} entry. In HX and SCD RBCs, CBD attenuates PIEZO1-TMEM16F coupling, thereby reducing PS exposure, microparticle release, thrombin generation, RBC-endothelium adhesion, and sickling. Beyond RBCs, CBD also blocks PIEZO2 currents and PIEZO2-dependent mechanical sensation in mice, suggesting broader effects of CBD-mediated PIEZO inhibition on nociceptive functions. Together, our findings identify CBD as a potent PIEZO inhibitor that restores calcium and membrane homeostasis, supporting the repurposing of CBD or the development of CBD-derived, PIEZO-selective analogs as a promising disease-modifying strategy for SCD, HX, and other PIEZO-mediated mechanosensing disorders. HighlightsO_LICBD inhibits PIEZO channels and disrupts the PIEZO1-TMEM16F axis in diseased RBCs C_LIO_LICBD shows a therapeutic window to prevent PS exposure and translational promise for HX and SCD C_LI

pharmacology and toxicology↗

bioRxiv2025

Population-specific brain charts reveal Chinese-Western differences in neurodevelopmental trajectories

Human brain charts provide unprecedented opportunities for decoding neurodevelopmental milestones and establishing clinical benchmarks for precision brain medicine 1-7. However, current lifespan brain charts are primarily derived from European and North American cohorts, with Asian populations severely underrepresented. Here, we present the first population-specific brain charts for China, developed through the Chinese Lifespan Brain Mapping Consortium (Phase I) using neuroimaging data from 43,037 participants (aged 0-100 years) across 384 sites nationwide. We establish the lifespan normative trajectories for 296 structural brain phenotypes, encompassing global, subcortical, and cortical measures. Cross-population comparisons with Western brain charts (based on data from 56,339 participants aged 0-100 years) reveal distinct neurodevelopmental patterns in the Chinese population, including prolonged cortical and subcortical maturation, accelerated cerebellar growth, and earlier development of sensorimotor regions relative to paralimbic regions. Crucially, these Chinese-specific charts outperform Western-derived models in predicting healthy brain phenotypes and detecting pathological deviations in Chinese clinical cohorts. These findings highlight the urgent need for diverse, population-representative brain charts to advance equitable precision neuroscience and improve clinical validity across populations.

neuroscience↗

bioRxiv2025

Targeting PIEZO1-TMEM16F Coupling to Mitigate Sickle Cell Disease Complications

A deeper understanding of sickle cell disease (SCD) pathophysiology is critical for identifying novel therapeutic targets. A hallmark of SCD is abnormal phosphatidylserine (PS) exposure on sickle red blood cells (RBCs), which contributes to anemia, thrombosis, and vaso-occlusive crises (VOC). However, the mechanisms underlying this excessive PS exposure remain unclear. Here, we identify TMEM16F, a Ca2+-activated lipid scramblase, as a key mediator of PS exposure downstream of Ca2+ influx through the mechanosensitive channel PIEZO1 in sickle RBCs. Electrophysiology, imaging and flow cytometry reveal that deoxygenation-induced sickling promotes PIEZO1 activation, triggering Ca2+ entry, TMEM16F activation, and PS exposure. This cascade enhances PS+ microparticle release, thrombin generation, and RBC adhesion to endothelial cells. Notably, partial PIEZO1 inhibition with benzbromarone, an anti-gout drug, suppresses these changes. Our findings thus define a previously unrecognized mechanotransduction pathway in sickle RBCs and propose a unique therapeutic strategy to mitigate hypercoagulability and vaso-occlusion associated with SCD. Brief SummaryEnhanced PIEZO1 activation in sickle red blood cells promotes TMEM16F scramblase-mediated phosphatidylserine exposure and subsequent sickle cell disease complications. Disrupting this coupling presents a potential therapeutic strategy.

physiology↗

bioRxiv2025

RAD51AP1 is a versatile RAD51 modulator

RAD51AP1 is an emergent key factor in homologous recombination (HR), the major pathway for accurate repair of DNA double-strand breaks, and in alternative lengthening of telomeres (ALT). Depletion of RAD51AP1 diminishes HR and overexpression is common in cancer, where it is associated with malignancy. Here, we show that RAD51AP1 serves as a versatile modulator of the RAD51 recombinase, the central player in HR. Through a combination of biochemistry and structural biology, we reveal that RAD51AP1 possesses at least three RAD51-binding sites that facilitate its binding across two adjacent RAD51 molecules. We uncover a novel RAD51 binding mode and characterise a hitherto overlooked role for RAD51AP1 in stabilising RAD51-ssDNA filaments and promoting strand exchange. Further, we resolved structures of RAD51-ssDNA filaments in the presence of Mg2+-ATP and Mg2+-ADP, revealing conformational changes upon ATP hydrolysis and explaining how ADP reduces RAD51-DNA binding. Our findings provide mechanistic insights into RAD51 recombinase and RAD51AP1. HighlightsO_LIStructures of RAD51 filaments in the presence of Mg2+-ATP and Mg2+-ADP reveal how ATP hydrolysis reduces DNA binding C_LIO_LIATP hydrolysis does not induce RAD51 filament contraction, instead it induces filament relaxation C_LIO_LIRAD51AP1 uses three sites to bind across two RAD51 monomers C_LIO_LIRAD51AP1 uses a unique binding mode to stabilise the RAD51 N-terminal domain and protomer interface of RAD51 C_LIO_LIRAD51AP1 binding induces conformational changes that promote RAD51 DNA association, oligomerisation, filament nucleation, filament stabilisation and strand exchange C_LI

biochemistry↗

bioRxiv2025

Calpain cleaves the carboxyl terminus of TRPV1 and modulates receptor tachyphylaxis

Transient receptor potential vanilloid-1 (TRPV1) plays a critical role in noxious heat sensation and pain hypersensitivity development in chronic pain. Sustained or repeated exposure to capsaicin, a classic TRPV1 agonist, induces TRPV1 desensitization. This partially accounts for the analgesic effect of capsaicin. However, the regulatory mechanisms of TRPV1 desensitization remain poorly understood. In this study, we found that capsaicin acts on TRPV1 to induce the activation of calpain, a Ca2+-sensitive cysteine protease, in a manner unrelated to cellular injury. Calpain cleaves rTRPV1 at the carboxyl terminus of G819/S820. Lack of the distal carboxyl terminus leads to reduced TRPV1 localization on the plasma membrane, potentially due to increased receptor internalization and impaired subunit assembly. This finding was corroborated by whole-cell patch clamp recordings. Additionally, the {Delta}820-838 mutant of rTRPV1 shows resistance to tachyphylaxis as induced by repetitive capsaicin stimulation. This study reveals a pivotal role for calpain in TRPV1 desensitization where its activation constrains TRPV1 channel function while simultaneously increasing its resistance to tachyphylaxis, thereby acting to maintain TRPV1 activity within an appropriate range.

neuroscience↗

bioRxiv2025

PIEZO1 Drives Trophoblast Fusion and Placental Development

PIEZO1, a mechanosensor1,2 in endothelial cells, plays a critical role in fetal vascular development during embryogenesis3,4. However, its expression and function in placental trophoblasts remain unexplored. Here, we demonstrate that PIEZO1 is expressed in placental villus trophoblasts, where it is essential for trophoblast fusion and placental development. Mice with trophoblast-specific PIEZO1 knockout exhibit embryonic lethality without obvious vascular defects. Instead, PIEZO1 deficiency disrupts the formation of the syncytiotrophoblast layer in the placenta. Mechanistically, PIEZO1-mediated calcium influx activates TMEM16F lipid scramblase, facilitating the externalization of phosphatidylserine, a key "fuse-me" signal for trophoblast fusion5,6. These findings reveal PIEZO1 as a crucial mechanosensor in trophoblasts and highlight its indispensable role in trophoblast fusion and placental development, expanding our understanding of PIEZO1s functions beyond endothelial cells during pregnancy.

cell biology↗

bioRxiv2024

SegCSR: Weakly-Supervised Cortical Surfaces Reconstruction from Brain Ribbon Segmentations

Deep learning-based cortical surface reconstruction (CSR) methods heavily rely on pseudo ground truth (pGT) generated by conventional CSR pipelines as supervision, leading to dataset-specific challenges and lengthy training data preparation. We propose a new approach for reconstructing multiple cortical surfaces using weak supervision from brain MRI ribbon segmentations. Our approach initializes a midthickness surface and then deforms it inward and outward to form the inner (white matter) and outer (pial) cortical surfaces, respectively, by jointly learning diffeomorphic flows to align the surfaces with the boundaries of the cortical ribbon segmentation maps. Specifically, a boundary surface loss drives the initialization surface to the target inner and outer boundaries, and an inter-surface normal consistency loss regularizes the pial surface in challenging deep cortical sulci. Additional regularization terms are utilized to enforce surface smoothness and topology. Evaluated on two large-scale brain MRI datasets, our weakly-supervised method achieves comparable or superior CSR accuracy and regularity to existing supervised deep learning alternatives.

neuroscience↗

bioRxiv2024

Nanoparticles use magnetoelectricity to target and eradicate cancer cells

This study presents the first in vivo and in vitro evidence of an externally controlled, predictive, MRI-based nanotheranostic agent capable of cancer cell specific targeting and killing via irreversible electroporation (IRE) in solid tumors. The rectangular-prism-shaped magnetoelectric nanoparticle is a smart nanoparticle that produces a local electric field in response to an externally applied magnetic field. When externally activated, MENPs are preferentially attracted to the highly conductive cancer cell membranes, which occurs in cancer cells because of dysregulated ion flux across their membranes. In a pancreatic adenocarcinoma murine model, MENPs activated by external magnetic fields during magnetic resonance imaging (MRI) resulted in a mean three-fold tumor volume reduction (62.3% vs 188.7%; P < .001) from a single treatment. In a longitudinal confirmatory study, 35% of mice treated with activated MENPs achieved a durable complete response for 14 weeks after one treatment. The degree of tumor volume reduction correlated with a decrease in MRI T2* relaxation time (r = .351; P = .039) which suggests that MENPs have a potential to serve as a predictive nanotheranostic agent at time of treatment. There were no discernable toxicities associated with MENPs at any timepoint or on histopathological analysis of major organs. MENPs are a noninvasive alternative modality for the treatment of cancer. SummaryWe investigated the theranostic capabilities of magnetoelectric nanoparticles (MENPs) combined with MRI via a murine model of pancreatic adenocarcinoma. MENPs leverage the magnetoelectric effect to convert an applied magnetic field into local electric fields, which can induce irreversible electroporation of tumor cell membranes when activated by MRI. Additionally, MENPs modulate MRI relaxivity, which can be used to predict the degree of tumor ablation. Through a pilot study (n=21) and a confirmatory study (n=27), we demonstrated that, [≥]300 {micro}g of MRI-activated MENPs significantly reduced tumor volumes, averaging a three-fold decrease as compared to controls. Furthermore, there was a direct correlation between the reduction in tumor T2 relaxation times and tumor volume reduction, highlighting the predictive prognostic value of MENPs. Six of 17 mice in the confirmatory studys experimental arms achieved a durable complete response, showcasing the potential for durable treatment outcomes. Importantly, the administration of MENPs was not associated with any evident toxicities. This study presents the first in vivo evidence of an externally controlled, MRI-based, theranostic agent that effectively targets and treats solid tumors via irreversible electroporation while sparing normal tissues, offering a new and promising approach to cancer therapy.

bioengineering↗

bioRxiv2024

DEK-rRNA interactions regulate ribosome biogenesis and stress response

DNA/chromatin-based functions of the DEK oncogene, a unique nucleic acid-interacting factor in metazoans, have been widely investigated, yet its role in cellular RNA biology is under-studied. Herein we employed CLIP-seq alongside mutational, biochemical, and cellular approaches to gain deeper insights into the cellular DEK-RNA interplay. We confirm interaction of DEK with coding RNA, yet also report association with ribosomal RNA (rRNA) and thereby establishing a robust link between DEK and ribosome biology. Indeed, cells lacking DEK showed marked deficits in cytoplasmic ribosome quality and function. This phenotype was exclusively rescued by C-terminal DEK, harboring two RNA interaction domains, but not by an rRNA-binding deficient mutant. Mechanistically, we uncovered pleiotropic involvement of DEK in RNA polymerase I-mediated rRNA transcription and processing pathways. More specifically, we found direct interaction of DEK with RNA polymerase III-transcribed 5S rRNA and identified DEK as a regulator of the Impaired Ribosome Biogenesis Checkpoint (IRBC). Within this ribosomal stress pathway, DEK depletion results in free 5S RNP, triggering stabilization of p53 via inhibition of MDM2. In summary, our multilayer analysis revealed DEK as a potent cellular RNA binding protein and provides first evidence of DEK as a regulator of ribosome biogenesis and stress response via the 5S RNP-MDM2-p53 axis.

biochemistry↗

bioRxiv2024

AmygdalaGo-BOLT3D: A boundary learning transformer for tracing human amygdala

Each year, thousands of brain MRI scans are collected to study structural development in children and adolescents. However, the amygdala, a particularly small and complex structure, remains difficult to segment reliably, especially in developing populations where its volume is even smaller. To address this challenge, we developed AmygdalaGo-BOLT, a boundary-aware deep learning model tailored for human amygdala segmentation. It was trained and validated using 854 manually labeled scans from pediatric datasets, with independent samples used to ensure performance generalizability. The model integrates multiscale image features, spatial priors, and self-attention mechanisms within a compact encoder-decoder architecture to enhance boundary detection. Validation across multiple imaging centers and age groups shows that AmygdalaGo-BOLT closely matches expert manual labels, improves processing efficiency, and outperforms existing tools in accuracy. This enables robust and scalable analysis of amygdala morphology in developmental neuroimaging studies where manual tracing is impractical. To support open and reproducible science, we publicly release both the labeled datasets and the full source code. TeaserAn AI model enables accurate amygdala segmentation across ages and centers, supporting large-scale brain imaging studies.

neuroscience↗

bioRxiv2024

Enhancing Whole Slide Image Classification with Discriminative and Contrastive Learning

Whole slide image (WSI) classification plays a crucial role in digital pathology data analysis. However, the immense size of WSIs and the absence of fine-grained sub-region labels, such as patches, pose significant challenges for accurate WSI classification. Typical classification-driven deep learning methods often struggle to generate compact image representations, which can compromise the robustness of WSI classification. In this study, we address this challenge by incorporating both discriminative and contrastive learning techniques for WSI classification. Different from the extant contrastive learning methods for WSI classification that primarily assign pseudo labels to patches based on the WSI-level labels, our approach takes a different route to directly focus on constructing positive and negative samples at the WSI-level. Specifically, we select a subset of representative and informative patches to represent WSIs and create positive and negative samples at the WSI-level, allowing us to better capture WSI-level information and increase the likelihood of effectively learning informative features. Experimental results on two datasets and ablation studies have demonstrated that our method significantly improved the WSI classification performance compared to state-of-the-art deep learning methods and enabled learning of informative features that promoted robustness of the WSI classification.

pathology↗

bioRxiv2024

TMEM16 and TMEM63/OSCA proteins share a conserved potential to permeate ions and phospholipids

The calcium-activated TMEM16 proteins and the mechanosensitive/osmolarity-activated OSCA/TMEM63 proteins belong to the Transmembrane Channel/Scramblase (TCS) superfamily. Within the superfamily, OSCA/TMEM63 proteins, as well as TMEM16A and TMEM16B, are thought to function solely as ion channels. However, most TMEM16 members, including TMEM16F, maintain an additional function as scramblases, rapidly exchanging phospholipids between leaflets of the membrane. Although recent studies have advanced our understanding of TCS structure-function relationships, the molecular determinants of TCS ion and lipid permeation remain unclear. Here we show that single mutations along the transmembrane helix (TM) 4/6 interface allow non-scrambling TCS members to permeate phospholipids. In particular, this study highlights the key role of TM 4 in controlling TCS ion and lipid permeation and offers novel insights into the evolution of the TCS superfamily, suggesting that, like TMEM16s, the OSCA/TMEM63 family maintains a conserved potential to permeate ions and phospholipids.

biophysics↗

bioRxiv2023

Long noncoding RNA Hottip maintained skeletal homeostasis via suppressing the enhancer of zeste homolog 2 (Ezh2)/histone methylation regulatory axis

Recent evidence suggests that long noncoding RNAs (lncRNAs) play a crucial role in regulating bone remodeling and skeletal homeostasis by coordinating the development of osteoblasts and osteoclasts. Several lncRNAs have been identified to participate in bone formation and resorption processes. Among them, the well-known oncogenic lncRNA, Hottip, has been reported to be involved in osteogenesis regulation. However, the specific function and underlying mechanisms remain poorly understood. In this study, we investigated the role of lncRNA Hottip in bone remodeling and skeletal homeostasis. Hottip knockout mice exhibited disrupted bone metabolism, abnormal bone tissue, and compromised bone quality, resulting in delayed fracture healing. In vitro experiments demonstrated that Hottip knockdown inhibited osteoblast differentiation while promoting osteoclast differentiation, with the opposite effect observed upon Hottip overexpression. Mechanistically, Hottip physically interacted with EZH2, leading to its degradation and facilitating the transcription of osteogenic genes by suppressing H3K9me3 and H3K27me3. In vivo experiments further validated the potential of Hottip overexpression to promote bone regeneration and accelerate fracture healing. In conclusion, our study reveals Hottip as a critical regulator in the differentiation of osteoblasts and osteoclasts, crucial for maintaining skeletal homeostasis. This lncRNA shows potential as a promising therapeutic target for bone regeneration.

cell biology↗