Biology subjects

Chang, J.

Publications and source records attributed to Chang, J..

At least 19 recordsLinked to original sources

bioRxiv2026

FMR1 Perturbations Disrupt Sensory Corticostriatal Circuits and Mitochondrial Energetics in Fragile X Syndrome

Fragile X syndrome (FXS) features sensory, social, and cognitive dysfunction, but how fragile X messenger ribonucleoprotein 1 (FMR1) deficiency disrupts sensory corticostriatal circuitry and whether distinct FMR1 perturbations converge on shared cellular mechanisms remain incompletely defined. We integrated diffusion magnetic resonance imaging (MRI) in individuals with FXS and Fmr1-knockout (KO) mice with projection-defined calcium recording, bidirectional optogenetic manipulation, primary somatosensory cortex (S1)-targeted expression of three patient-associated FMR1 missense variants, regional transcriptomics, and optical manipulation of mitochondrial energetic state. Individuals with FXS exhibited disrupted sensory corticostriatal structural connectivity, with mouse analyses further localizing abnormalities to S1-striatal pathways. Fmr1 deficiency impaired recruitment of the S1-dorsomedial striatum (DMS) pathway during social interaction, and projection-specific manipulation demonstrated that S1-DMS dysfunction contributes to social and cognitive deficits. Three patient-associated FMR1 variants, I234T, E329G, and R586Q, converged on social behavioral deficits, which were improved by S1-DMS activation. Transcriptomic analyses revealed convergent oxidative phosphorylation (OXPHOS)-related mitochondrial remodeling in S1 and DMS across Fmr1 deficiency and variant models. Systemic treatment with the mitochondria-targeted antioxidant MitoQ attenuated novel-location-recognition, social-interaction, and social-novelty deficits in Fmr1-KO mice. Moreover, mt-EcGAPR-based Mitochondrial OptoEnergetic Activation (Mito-OptoEA) enhanced mitochondrial energetic state in S1 and improved novel-location recognition and sociability in both Fmr1-KO and I234T models. Together, these findings define a sensory corticostriatal circuit-metabolic framework for FXS in which S1-DMS circuit dysfunction and convergent mitochondrial energetic abnormalities contribute to FMR1-related social and cognitive phenotypes.

neuroscience↗

bioRxiv2026

Lipid acyl chain remodeling governs KRAS nanoclustering and oncogenic functions through membrane composition control

KRAS signaling is spatially organized within the dynamic proteolipid nanoclusters on the plasma membrane (PM) that selectively concentrate anionic phospholipids, yet the molecular determinants governing this lipid sorting remain incompletely defined. Here we identify phospholipid acyl chain composition as a critical regulator of KRAS spatial organization and signal output. Using genetic and pharmacological perturbation of lysophosphatidylcholine acyltransferase 3 (LPCAT3), a key enzyme that incorporates polyunsaturated fatty acids into phospholipids, we demonstrate that remodeling of membrane acyl chain unsaturation alters the phosphatidylserine (PS) lipidome without changing total PS abundance. Loss of LPCAT3 depletes polyunsaturated PS species, disrupts KRAS PM association and nanoclustering, and attenuates downstream MAPK signaling and oncogenic activity across multiple KRAS mutant contexts. Acute restoration of polyunsaturated, but not saturated, lipids rescue KRAS nanoclustering, establishing a direct requirement for lipid unsaturation in sustaining signaling-competent assemblies. Mechanistically, these findings define membrane acyl chain composition as a determinant of proteolipid organization that encodes KRAS signaling capacity. Consistent with this model, LPCAT3 inhibition selectively impairs KRAS-dependent proliferation and sensitizes tumor cells to direct KRAS inhibition. Together, this work uncovers lipid acyl chain remodeling as a fundamental regulator of membrane-associated signaling and identifies membrane composition as an exploitable vulnerability in oncogenic KRAS systems.

cancer biology↗

bioRxiv2026

Specification dependence of EEG theta/alpha ratio associations with cognitive impairment: a multiverse analysis

Objective: The theta/alpha ratio, an index of electroencephalographic slowing, has been proposed as a marker of cognitive impairment, but estimates may depend on preprocessing, spectral, spatial, and statistical decisions. We used multiverse analysis to evaluate its robustness across plausible pipelines. Methods: Two resting-state EEG datasets were analyzed. Dataset 1 included 49 controls and 100 participants with Parkinson's disease spanning normal cognition to dementia. Dataset 2 included 29 controls, 36 patients with Alzheimer's disease, and 23 with frontotemporal dementia. Eight decisions were varied: filtering, artifact correction, independent component analysis (ICA), referencing, spatial summary, alpha-band definition, spectral method, and covariate adjustment, yielding 1,280 specifications (universes). Results: Findings varied across the unrestricted multiverse. In Dataset 1, all universes in the exploratory family combining ICA with absolute fast Fourier transform (FFT) or Welch power yielded p < 0.05 for comparisons of controls and cognitively normal Parkinson's disease with Parkinson's disease dementia and produced significant associations with Montreal Cognitive Assessment scores in more than 95% of universes. In Dataset 2, absolute FFT or Welch power supported control versus Alzheimer's disease differences in more than 95% of universes, while ICA with absolute power yielded significant Mini-Mental State Examination associations in all specifications. Higher theta/alpha ratios were consistently associated with greater cognitive impairment. Conclusions: The theta/alpha ratio is a promising but specification-dependent marker of cognitive impairment, contingent particularly on ICA and spectral power quantification. Significance: Identifying the analytic conditions under which theta/alpha findings remain stable supports standardized, reproducible evaluation of quantitative EEG markers of cognitive impairment.

neuroscience↗

bioRxiv2026

ApoE4 impairs astrocyte lipid droplet formation via endolysosomal dysfunction

An important component of neuron-glia coupling is the transport and delivery of neuron-derived lipids to glial lipid droplets. This pathway is important as failure to store incoming lipids in glial lipid droplets exacerbates neurodegeneration. ApoE4, a risk factor for Alzheimer's disease, impairs this transport pathway. But how ApoE4 affects lipid trafficking in glia and whether these alterations can be restored is poorly understood. Here, we demonstrate that ApoE4 particles impair endolysosomal function and promote lipofuscin formation in cultured primary astrocytes, thereby disrupting the trafficking of lipids for storage into lipid droplets. Lipid droplets, however, can be recovered by preventing lysosomal impairment with PCSK9, a secreted protein that prevents LDLR recycling and reduces ApoE4 internalization. Lipid droplets are similarly recovered in the presence of ApoE4 by repairing endolysosomal function. Our findings reveal new insight into how ApoE4 dysregulates lipid storage while uncovering new mechanisms to correct these defects.

neuroscience↗

bioRxiv2026

Optineurin Deficiency Collapses the Host Endolysosomal Network and Impairs Xenophagy to Accelerate Mycobacterium tuberculosis Growth

Selective autophagy is a host defense mechanism against Mycobacterium tuberculosis (Mtb) that restricts bacterial growth by targeting ubiquitin-coated bacilli for lysosomal degradation via autophagy receptors. Optineurin is a selective autophagy receptor that targets pathogens and modulates immune signaling; however, its precise structural mechanism during Mtb infection remains poorly defined. Here, we show that while Optineurin deficiency spares the global host transcriptomic response to infection, it collapses the host endolysosomal network, reducing LAMP1+ and LysoTracker+ reserves by half. Multi-dose bafilomycin A1 flux assays demonstrated that this structural depletion selectively blocks the dynamic, directional trafficking and functional delivery of autophagosomes to the pathogen, significantly reducing Mtb-DQ-BSA colocalization. Genetic complementation restored bacterial restriction in a manner dependent on three phosphosites (Ser187, Ser530, and the uncharacterized Ser556). In the context of reduced autophagic containment and increased Mtb replication, Optineurin deficiency accelerated necrotic-like host cell death. In vivo, Optineurin deficiency enhanced bacterial replication and impaired the Type I interferon response during acute Mtb infection but did not affect long-term survival. Together, these findings identify Optineurin as a critical regulator of autophagic flux, host cell death, and Type I interferon responses that limit early Mtb pathogenesis.

microbiology↗

bioRxiv2026

De novo design of small-molecule-induced conformational change

Many biological proteins function by changing shape upon small-molecule binding. Here, we present a general strategy for designing de novo proteins that undergo small-molecule-induced conformational change. Our approach converts a preorganized small-molecule binding protein into a ligand-responsive shape-changer by adding a mobile lid domain that closes behind the ligand upon binding. Using this strategy, we converted an exatecan-binding protein into a drug-induced conformational switch. The lidded proteins showed considerably stronger binding affinity in the sub-nanomolar regime, 100-fold greater specificity to exatecan over a similar molecule, and ligand residence times up to several months, with tunable binding kinetics. We turned one design into a genetically encodable fluorescent biosensor of the drug, enabling potential clinical applications. Our results open the door to programming complex molecular function using vast chemical space.

bioengineering↗

bioRxiv2026

Targeting cellular senescence alleviates bone marrow aging

Aging of the hematopoietic system impairs hematopoietic stem cell (HSC) function and alters bone marrow niche behavior, increasing susceptibility to anemia, infections, and hematologic malignancies. Here, pharmacologic clearance of senescent cells with the PROTAC compound 753b simultaneously targeting BCL-xL and BCL-2 reverses key secretory, transcriptional, and functional hallmarks of hematopoietic aging with low toxicity, restoring balanced lineage output. Single-cell RNA sequencing further demonstrates that 753b treatment attenuates aging-associated transcriptional signatures in HSCs, while selectively eliminating senescent, pro-survival niche cells without grossly perturbing niche composition. Functionally, 753b suppresses pro-inflammatory cues from both niche and hematopoietic cells including those emanating from neutrophil progenitors, rebalancing global bone marrow secretory ecosystem across stromal and hematopoietic compartments. Collectively, we identify 753b-induced senescent cell clearance as a powerful strategy to rejuvenate aged hematopoiesis and re-establish homeostatic communication between HSCs and their microenvironment, with implications for mitigating age-related hematologic dysfunction and improving hematologic health in older individuals.

cell biology↗

bioRxiv2026

Human meningiomas arise from developmental meningeal mosaicism

Meningioma is the most common primary intracranial tumour, yet its genetic origin and the temporal sequence of mutational events remain poorly defined. Here, we analysed 80 triple-matched tumour, histologically normal meninges, and blood samples from 22 patients. NF2 or TRAF7 driver mutations are detectable in phenotypically normal meninges in 81.8% of cases (95% CI: 59.7-94.8%; VAF [~]0.02%), corroborated by high-depth sequencing, single-cell cloning, and phylogenetic analysis. By distinguishing developmental mosaic mutations from postnatal tumour-private mutations, we revealed distinct mutational signatures and resolved the temporal sequence of meningioma evolution. The developmental origin was further underscored in patients with multiple meningiomas, where identical driver mutations were shared across genomically distinct tumours and meninges, and in intraventricular meningiomas (IVM), where driver mutations were detected in distant cranial dura. Reconstruction of mutational timing revealed lineage-specific trajectories, linking mosaicism to diverse disease presentations including solitary meningioma, IVM, meningiomatosis, and NF2-related schwannomatosis. Together, these findings reveal an early origin of human meningioma, in which developmental mosaicism establishes a pre-neoplastic field within the meninges, providing a developmental framework for adult tumourigenesis.

Cancer Biology↗

bioRxiv2026

Trichome and inflorescence evolution in the paleopolyploid tree genus Greyia Hook. & Harv.

The southern African endemic tree genus Greyia Hook. & Harv. (Francoaceae, Geraniales) comprises three species distinguished by striking variation in trichome morphology and floral architecture, yet the genomic and regulatory basis of these traits has remained largely unexplored. Here, we present chromosome-scale genome assemblies for all recognized Greyia species, namely Greyia radlkoferi, Greyia sutherlandii, and Greyia flanaganii, providing the first high-quality genomic resources for Geraniales. Phylogenomic analyses placed Geraniales as sister to Crossosomatales, with divergence at [~]103 million years ago (Mya), and indicated that Greyia diversified recently during the Quaternary ([~]1.39 Mya). Comparative genomics revealed a Greyia-specific whole-genome triplication event dated to [~]86.9 Mya, generating extensive gene duplication which has been retained. Transcriptomic analyses linked divergence in trichome morphology to regulatory subfunctionalization of the conserved MYB-bHLH-WD40 (MBW) complex, including transcriptional silencing of GL1 and shifts in GL3/EGL3 expression in G. sutherlandii. In addition, analyses of flower and inflorescence identified expansion of MADS-box transcription factor families and species-specific cis-regulatory divergence in LFY and FT. Notably, predicted AGL6 binding sites are retained in the LFY and FT promoters of G. radlkoferi and G. sutherlandii but absent in G. flanaganii, which aligns with differences in inflorescence density among species. Together, these results demonstrate how polyploidy and regulatory divergence of conserved developmental pathways have shaped morphological diversification in Greyia. Significance StatementUnderstanding how morphological diversity arises from conserved pathways is a central evolutionary challenge, particularly in lineages shaped by ancient polyploidy. We present the first chromosome-scale genomes for the order Geraniales, focusing on three southern African endemic Greyia species. We demonstrate that regulatory divergence following a lineage-specific whole-genome triplication, rather than changes in gene content, drives variation in trichome morphology and floral architecture.

genomics↗

bioRxiv2026

Sphk1-S1P signaling drives blood-brain barrier breakdown after intracerebral hemorrhage via HIF-1α-dependent upregulation of Bsg-MMP-9

Blood-brain barrier (BBB) breakdown is a critical pathological event driving secondary brain injury and poor outcomes following intracerebral hemorrhage (ICH). However, the mechanisms governing acute BBB breakdown after ICH remain incompletely understood. Here we demonstrate that the Sphk1-S1P-S1PR3 signaling plays a pivotal role in this process. Sphk1 expression was significantly upregulated in the perihematomal endothelial cells of both ICH patients and mice, with levels positively correlating with BBB dysfunction severity and poor clinical outcomes. Using endothelial-specific genetic gain- and loss-of-function approaches, we found that Sphk1 knockdown attenuated BBB leakage, reduced hematoma volume and brain edema, preserved tight junction integrity, and improved neurological function at 1-day post-ICH, whereas Sphk1 overexpression exacerbated these pathological features. Mechanistically, transcriptomic profiling of perihematomal endothelial cells revealed that prior to its established role in Nlrp3-mediated pyroptosis, Sphk1 promotes early BBB breakdown by regulating the Bsg-MMP-9 axis. Endothelial-specific Bsg deletion completely abrogated the deleterious effects of Sphk1, confirming Bsg as an indispensable intermediary through which Sphk1 signals to MMP-9. ATAC-seq and dual-luciferase assays further demonstrated that Sphk1-generated S1P signals through S1PR3 to activate HIF-1, which directly binds the Bsg promoter to drive its transcription, ultimately promoting MMP-9-mediated tight junction degradation. These findings delineate a complete hierarchical signaling cascade from metabolic enzyme to transcriptional regulation and subsequent barrier injury, establishing the Sphk1-Bsg-MMP-9 axis as a promising therapeutic target for ICH. One sentence summaryThis work identifies an early and pivotal mechanism of blood-brain barrier breakdown after intracerebral hemorrhage, demonstrating that Sphk1-generated S1P signals through S1PR3 to activate HIF-1, which directly transactivates Bsg expression, leading to MMP-9-mediated tight junction degradation, thereby establishing a novel hierarchical axis with therapeutic potential.

cell biology↗

bioRxiv2026

Loss of lmx1ba drives premature osteoarthritis through disruption of skeletal homeostasis

Osteoarthritis is increasingly recognised as a disease of failed integration across the whole joint unit; however, the mechanisms that co-ordinate tissue integrity from development through to adult homeostasis remain largely unresolved. The LIM-homeodomain transcription factor LMX1B is a key determinant of embryonic skeletal patterning, but how it functions to regulate skeletal integrity in the mature skeleton is unknown. Recently, LMX1B was identified as a key driver of osteoarthritis. Here we show that loss of lmx1ba in zebrafish causes premature and progressive severe osteoarthritic pathology in adult spines and jaws, despite largely normal early skeletal patterning, revealing a conserved and continuous requirement for lmx1ba in joint maintenance beyond development. At a cellular level, loss of lmx1ba decouples osteoblast and osteoclast-mediated remodelling leading to bone overgrowth, heterogeneity of bone properties causing increased incidence of spontaneous fractures, and progressive abnormalities in spine morphology. In parallel, we observe degeneration of the intervertebral disc and dysregulation of the proteome and glycosaminoglycans indicative of disrupted extracellular matrix and a breakdown of the coordinated regulation of hard and soft tissue interfaces, which at the organismal level leads to altered joint performance. Notably, degeneration is restricted to mobile joints, and is not observed in cranial sutures, demonstrating a selective requirement for lmx1ba in mechanically active tissues. These changes are consistent with a model of spatially disrupted matrix properties that, under cyclic loading, promote progressive tissue damage. Our findings support a model in which continued expression of LMX1B in adulthood is required to maintain joint structures throughout life.

physiology↗

bioRxiv2026

Generation and long-term expansion of human pancreatic islet organoids in vitro

The scarcity of expandable, functional human islet cells remains a major barrier to diabetes therapy. Here, we identify PROCR+ cells within adult human islets and establish a defined culture system to generate pancreatic islet organoids. These organoids self-organize into -, {beta}-, {delta}-, and PP cells at near-native ratios, exhibit regulated insulin and glucagon secretion, and support exponential in vitro expansion. Single-cell transcriptomics reveals a unique progenitor-like cell population that is transcriptionally primed for endocrine differentiation but shares molecular features with fetal trunk cells and endocrine progenitors. When transplanted, the organoids rapidly ameliorate hyperglycemia in diabetic mice. Importantly, in a non-human primate model, intraportal transplantation of these organoids reduced exogenous insulin requirements, restored glucose-stimulated C-peptide secretion, and achieved sustained glycemic control-representing a critical step toward clinical translation. This study provides a strategy for expanding human islet organoids, offering a scalable platform for diabetes treatment, disease modeling, and regenerative medicine.

cell biology↗

bioRxiv2026

Transcription Factor Subtype Governs Response and Resistance to DLL3-Directed T-Cell Engagement in Small Cell Lung Cancer

Although small cell lung cancer (SCLC) comprises transcription factor (TF)-defined molecular subtypes (ASCL1, NEUROD1, POU2F3), the extent to which these subtypes predict response to clinically effective therapy in patients--and whether therapy can select for subtype switching--remains unknown. The recent approval of the DLL3xCD3 bispecific T-cell engager tarlatamab represents one of the first meaningful advances in relapsed small cell lung cancer (SCLC) in decades, yet responses remain heterogeneous and resistance is inevitable. Here, we inferred SCLC gene expression from circulating chromatin in prospectively collected patient plasma (46 patients; 167 samples), enabling interrogation of response and acquired resistance to tarlatamab. Parallel development of the first immunocompetent syngeneic mouse model to study tarlatamab response and resistance enabled functional validation. Across species, findings converged on a central principle: TF subtype governs both initial response and acquired resistance. Therapeutic response was significantly associated with ASCL1-subtype tumors, whereas NEUROD1-subtype tumors exhibited inferior responses and POU2F3-subtype tumors were uniformly resistant, consistent with DLL3 being a direct ASCL1 transcriptional target and most highly expressed in ASCL1-positive tumors. Strikingly, one mode of acquired resistance revealed therapeutic selection for a NEUROD1-high state with concomitant DLL3 downregulation. Other resistant tumors exhibited enrichment of regulatory and exhausted T-cell programs, highlighting tarlatamabs dual-targeting mechanism of action. Together, these results reveal that tarlatamab exerts selective pressure against ASCL1-driven lineages, facilitating resistance through loss of an antigen intrinsically linked to that state. These findings underscore the clinical relevance of TF-defined molecular subtypes in human SCLC. More broadly, they highlight the power of integrating longitudinal in vivo plasma transcriptional profiling from patient plasma with functional mouse modeling to uncover clinical and biological mechanisms of response and resistance to cell-surface-targeted therapies.

cancer biology↗

bioRxiv2026

Nextstrain automates real-time phylodynamic analysis of open data for endemic and emerging pathogens

MotivationGenome sequencing provides an exceptional window into the evolutionary and epidemiological dynamics of endemic and emerging pathogens, and thus allows for better, more targeted, public health interventions. Online genomic surveillance platforms can provide near real-time insight into these dynamics. ResultsNextstrain provides continually updated real-time genomic surveillance for 21 viruses and the bacterial pathogen Mycobacterium tuberculosis, with most analyses relying solely on open sequence data. Each pathogen includes steps to fetch and curate open data, classify sequences using established nomenclature systems, perform phylogenetic analyses, and share the results publicly. These analyses are automated, with most running daily to provide continually updated snapshots of pathogen evolution. Availability and ImplementationAll source code is available at https://github.com/nextstrain. Phylogenetic results can be visualized and downloaded at https://nextstrain.org/pathogens, and open sequence data and curated metadata are available at https://nextstrain.org/pathogens/files.

bioinformatics↗

bioRxiv2026

Thermoregulatory Constraints on Regenerative Competence: Evolutionary Trade-Offs Between Metabolic Homeostasis and Tissue Repair

Regenerative capacity varies widely across the animal kingdom, yet adult mammals and birds exhibit limited ability to regenerate complex tissues such as the central nervous system (CNS). The evolutionary basis for this loss of regenerative competence remains poorly understood. Here we advance a Thermoregulatory Theory of Regenerative Scope, proposing that the emergence and refinement of endothermic physiology shaped tissue-specific molecular architectures that can bias injury responses away from regeneration in highly metabolic organs. Rather than imposing a systemic energetic constraint, we suggest that endothermy provided the evolutionary context in which thermogenic calcium-handling systems became increasingly specialized in excitable tissues. In particular, futile Ca{superscript 2} cycling across the sarco/endoplasmic reticulum through SERCA pumps contributes to heat generation but may also promote sustained intracellular Ca{superscript 2} elevations that activate inflammatory and fibrotic/gliotic signaling pathways incompatible with functional regeneration. Comparative observations indicate that regenerative competence is retained in ectothermic vertebrates and in mammalian contexts where thermoregulatory systems are developmentally immature or physiologically attenuated. Importantly, regenerative outcomes differ substantially among tissues within endothermic organisms, suggesting that local calcium regulatory architectures, rather than metabolic intensity alone, may determine regenerative potential. This framework generates experimentally testable predictions linking thermogenic Ca{superscript 2} signaling to regenerative failure and provides an evolutionary lens through which disparate observations in regenerative biology may be unified.

evolutionary biology↗

bioRxiv2026

Ageing impacts extracellular matrix turnover and remodelling in the kidney

At least 10% of the global population is impacted by chronic kidney disease (CKD) and ageing is a key risk factor. CKD is characterised by the build-up of extracellular matrix and a loss of functional nephrons. However, the mechanisms that maintain matrix homeostasis across the physiological lifespan remain elusive. Using {superscript 1}3C-lysine metabolic labelling, we quantified kidney matrix protein turnover in healthy mice at four timepoints (8, 22, 52, and 78 weeks). We found that basement membrane components, including collagen IV, laminin-521, nidogens and perlecan, were more long-lived over age, with collagen IV half-lives extending from weeks in young kidneys to years in aged kidneys, suggesting a reduced capacity for basement membrane renewal. The half-lives of fibrillar collagens I and III also increased over age up to forty-fold, which is consistent with minimal degradation. In contrast, collagen XV retained rapid turnover despite increased abundance, indicating a persistent role in tissue remodelling. Using peptide location fingerprinting to predict structural alterations and proteolytic processing we identified age-dependent meprin oligomerisation and altered nidogen-laminin interaction states. We predicted structural alterations within assembly domains of collagen VI and reduced accessibility of integrin-binding regions, suggesting altered microfibril organisation and cell-surface binding. Collagen XV had predicted structural changes across the NC1 domain encoding the matrikine restin, consistent with altered protease accessibility and matrikine release during ageing. These findings indicate that age-related kidney fibrosis is primarily caused by impaired matrix degradation, with protease accessibility and altered matrix interactions likely playing key roles in this remodeling process.

systems biology↗

bioRxiv2026

Targeted therapy-induced chromosomal instability dictates mitotic dependency on Aurora Kinase A

Targeted therapies eliminate cancer cells by inhibiting oncogenic signaling; however, tumor cells often evade cytotoxicity through proteomic and epigenetic reprogramming that enables survival. These adaptive responses may create collateral cellular stresses, such as DNA damage, that can be therapeutically exploited. When unresolved, DNA damage leads to chromosomal instability (CIN), a potential source of vulnerability. Whether KRAS inhibition induces DNA damage or CIN in KRAS-mutant non-small cell lung cancer (NSCLC) has not been established. Here, we show that the KRASG12C inhibitor LY3499446 induces CIN in KRAS-mutant NSCLC cell lines. A targeted compound screen revealed that the extent of CIN induction by KRASG12C inhibition strongly correlates with therapeutic synergy with the selective Aurora kinase A inhibitor LSN3321213. Mechanistically, KRASG12C inhibition stabilizes cyclin B1 during mitosis through activation of mitotic ATR/ATM signaling. In the presence of Aurora Kinase A inhibition, cyclin B1 stabilization delays mitotic exit and diverts cell fate from mitotic slippage or division toward mitotic catastrophe. Together, our findings identify CIN as a predictive marker of response to combined KRASG12C and Aurora Kinase A inhibition, providing mechanistic rationale to enhance the therapeutic window of AURKA inhibitors when used with targeted therapies.

cancer biology↗

bioRxiv2025

Demonstrating and engineering the de novo synthesis of two aromatic chemicals estragole and anethole in Escherichia coli

Estragole and anethole are the major flavoring and aroma components of the essential oils extracted from basil (Ocimum basilicum L.), anise (Pimpinella anisum), star anise (Illicium verum), and fennel (Foeniculum vulgare). Besides, they also exhibit bioactivities, such as antioxidation, antimicrobial, immunomodulation, inti-inflammation, and anti-diabetes. These properties make them being faced with substantial applications in the industries of food, cosmetics, pharmaceutics, and agriculture. Commercial estragole and anethole are currently obtained from extraction from their original plants, which suffers from unstable quality and resources. Microbial synthesis is a promising alternative approach to sustainably obtain estragole and anethole, but has not been achieved. In this study, we modularly redesigned the synthetic pathways based on the Escherichia coli chassis. Firstly, a p-coumaryl alcohol over-accumulator (436.28 mg/L) was engineered to facilitate the validation of the undemonstrated downstream module. De novo biosynthesis of estragole and anethole was achieved by stepwise analysis and BioBricks assembly. The intracellular availability of S-adenosyl-L-methionine (SAM) was revealed to be the key limiting factor, and was enhanced by deregulating the synthesis. Finally, the productions of estragole and anethole were improved by 246.88% and 202.44% compared with the initial strains. This study represents the first synthesis of estragole and anethole in microbial chassis, and the demonstration of microbial synthesis of propenyl phenols. Besides, it provided an alternative sustainable approach to obtain flavoring and aroma chemicals.

synthetic biology↗