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Shi, Y.

Publications and source records attributed to Shi, Y..

At least 91 records · Page 5Linked to original sources

A Detailed Model for Understanding the Human Neocortex

The neocortex underlies cognitive abilities that set humans apart from other species. Although Ramon y Cajal initiated its study in the 19th century, much about its fundamental properties remain poorly understood. Biologically detail modeling, has been shown to serve as a tool to understand the modeled system better. By comparing computational models for different species we can highlight functional differences between them, find their anatomical or physiological basis and thus improve our understanding of cortical function. In this study we built a detailed model of a human cortical microcircuit following an established workflow. We compared the human data and results against a previously published reconstruction of rat cortical circuitry. To parametrize the human model, we gathered new original data on human morphological reconstructions, axonal bouton densities, single cell and synaptic recordings. We combined them with data available in the literature and open-sourced databases. We also developed various strategies to overcome the missing data, such as generalizing or adapting data from rodents. The resulting model consists of seven columnar units with similar characteristics. Each column has a radius of 476 {micro}m, a height of 2622 {micro}m, a volume of 1.86 mm3, a total cell density of 24,186 cells/mm3, on the order of 35,000 cells, around 12 million connections and approximately 47 million synapses. Comparing the rat and the human model showed that the human cortex is less dense in terms of cell bodies than the rodent cortex. Human cells have more complex branching, but lower bouton densities than rodent cells. However, the number of connections between cell types is similar.

neuroscience↗

NK Cells Engineered with a Chimeric Antigen Receptor Delay HIV Rebound and Reshape HIV Reservoir Composition

Durable HIV remission will require strategies that eliminate or permanently silence the latent reservoir that persists despite antiretroviral therapy (ART). Natural killer (NK) cells possess inherent antiviral activity, but unmodified NK cells have limited ability to recognize or clear latently infected cells during ART suppression. We engineered allogeneic human primary NK cells expressing a truncated CD4-based chimeric antigen receptor (D1D2-CAR) that targets the conserved CD4 binding site on HIV Env without permitting viral entry, and evaluated their activities in humanized mice infected with barcoded CCR5-tropic HIV. D1D2-CAR NK cells selectively killed HIV-infected primary CD4 T cells in vitro and significantly delayed viral rebound following ART interruption in humanized mice compared to GFP-NK or no NK control groups. Barcoded HIV tracking showed that CAR-NK treatment reduced the number, diversity, and inter-organ overlap of rebounding viral RNA and proviral DNA lineages, yielding rebound driven by a restricted set of dominant clones. Integration site and chromatin analysis further demonstrated selective depletion of proviruses positioned in genes, enhancers, promoters, and open chromatin. These findings show that CAR-NK cells can target rare reactivation events during ART suppression and reshape the reservoir toward a less rebound-competent, epigenetically repressive state.

immunology↗

SAGA/ATAC complexes sustain aberrant chromatin regulation and promote tumorigenesis in diffuse midline glioma

Diffuse midline gliomas (DMG) are aggressive pediatric brain tumors characterized by chromatin and transcriptional dysregulation induced by H3K27M mutations, with a median survival of 11-15 months. We identified multiple components of the SAGA and ATAC chromatin regulatory complexes as DMG genetic dependencies and found that genetic or pharmacological inhibition of the SAGA/ATAC-associated chromatin reader SGF29 reduces DMG proliferation in vitro and prolongs survival in xenograft models. Small molecule inhibitors targeting SAGA/ATAC-associated histone acetylation, ubiquitination, and methylation similarly suppress DMG growth. Integrative chromatin and transcriptomic profiling reveals that disruption of SAGA/ATAC through SGF29 knockout remodels the DMG chromatin landscape, producing distinct alterations at metabolic genes (loss of H3K9ac) and at embryonic/neurodevelopmental genes (redistribution of H3K4me3 and H3K27me3) and accompanying transcriptional changes. We further show that inhibition of the SAGA/ATAC-associated KAT2A/2B histone acetyltransferases represses cholesterol metabolism gene expression and that combined KAT2A/2B and cholesterol synthesis inhibition synergistically suppresses DMG growth in vitro and in a xenograft model. Together, these findings establish a mechanistic link between SAGA/ATAC-dependent chromatin regulation and the transcriptional and metabolic dysregulation underlying DMG malignancy. SignificanceWe demonstrate that SAGA/ATAC-dependent chromatin regulation sustains malignant transcription controlling cholesterol metabolism, proliferation, and cell fate in DMG, suggesting that a SAGA/ATAC-regulated epigenome-metabolome axis may be targeted to treat DMG.

cancer biology↗

Progranulin deficiency induces lipid droplet accumulation in microglia via a STAT3-GPAT3 axis

Heterozygous mutations in the progranulin (PGRN) encoding gene GRN cause frontotemporal dementia (FTD), whereas homozygous GRN mutations lead to neuronal ceroid lipofuscinosis (NCL). However, the mechanisms underlying neurodegeneration due to PGRN deficiency remain unclear. In the aged brains or under neurodegenerative conditions, the accumulation of lipid droplets (LDs) in microglia contributes to cellular dysfunction and pro-inflammatory responses, exacerbating neurodegenerative pathology. Here, we investigated how PGRN deficiency induces LD accumulation in microglia. We found that PGRN ablation significantly upregulated triglycerol-3-phosphate acyltransferase 3 (GPAT3), the first and rate-limiting enzyme for triacylglycerol biosynthesis, and increased LD formation in both Grn-/- BV2 microglial cells and Grn-/-mouse brains. Mechanistic study revealed that PGRN deficiency upregulates GPAT3 via stimulating signal transduction and activator of transcription factor 3 (STAT3) signaling. Silencing Gpat3, inhibiting GPAT3 activity with the small molecule inhibitor FSG67, or PGRN restoration reduced LD accumulation and rescued the cytotoxicity of Grn-/- microglia conditional medium (MCM) toward N2a cells. Notably, intraperitoneal administration of FSG67 or AAV-MG1.2-mCx3cr1-mGrn-eGFP-mediated microglia-specific gene therapy reduced microglial LDs and ameliorated behavior phenotypes in Grn-/- mice. Our study elucidates the role of PGRN loss in microglial lipid homeostasis and identifies the STAT3-GPAT3 axis as a potential therapeutic target for LD-associated neurodegenerative diseases.

neuroscience↗

Microhomology-mediated end joining acts directly on replication forks to repair single-ended double strand breaks

Replication stress, intrinsic to oncogenesis, often leads to fork breakage and double-strand break (DSB) formation. Conventionally, break-induced replication (BIR) is considered the primary mechanism for repairing replication-associated single-ended DSBs (seDSBs). Here, we demonstrate that microhomology-mediated end joining (MMEJ) acts directly to repair seDSBs at broken replication forks (fork-MMEJ), preferentially on the leading strands, and functions cooperatively with BIR. We also showed that while fork-MMEJ is promoted by Pol{theta}, it operates independently of MRE11/CtIP-mediated end resection, relies on RPA, and produces asymmetric deletion patterns, which is distinct from canonical MMEJ (cMMEJ) defined at replication-independent double-ended DSBs (deDSBs). ATR, activated as end resection proceeds, serves as a pivotal switch to suppress fork-MMEJ while promoting BIR. Combined inactivation of ATR and Pol{theta} synergistically kills cancer cells under high replication stress with minimal toxicity to normal cells. Together, our study provides fundamental insights into the MMEJ mechanism and offers new strategies for cancer treatment.

molecular biology↗

SoftHybrid: A Hybrid Imputation Algorithm Optimised for Single-Cell Proteomics Data

Missing values (MVs) remain a significant barrier to reliable proteomics analysis, particularly in single-cell proteomics, where small amounts of starting material and limits in detection drive Missing-Not-At-Random (MNAR) sparsity. Existing imputation methods typically target either Missing-At-Random (MAR) or MNAR mechanisms, resulting in a trade-off between replicate consistency and preservation of biological variation, and are largely designed for bulk data. Here, we introduce SoftHybrid, a data-driven imputation framework that jointly models missingness and protein abundance to estimate the probability of MNAR, enabling continuous weighting between MAR- and MNAR-oriented strategies. SoftHybrid requires no external priors (cell type labels, group annotations, predefined missingness assumptions, etc.), enabling fully unsupervised applications. Across ground truth benchmarks and real single-cell proteomics datasets, SoftHybrid outperforms existing methods at low input and matches or exceeds their performance at the mini-bulk level. By preserving proteomic structure and abundance accuracy, it enhances the recovery of biologically meaningful signals. SoftHybrid is implemented as an R package and is freely available on GitHub.

bioinformatics↗

EOLA1, a novel mitochondria-localized protein critical for heart functions via regulating mitochondrial translation

Mitochondria possess their own mRNA translation system, mediated by specialized mitoribosomes. Dysregulation of mitochondrial translation disrupts metabolic homeostasis and is linked to various pathologies, including metabolic syndromes, neurological disorders, and cardiac diseases. Using CRISPR/Cas9 screening, we identified EOLA1 (endothelial-overexpressed lipopolysaccharide-associated factor 1) as a novel mitochondrial matrix protein essential for metabolic homeostasis. Knockout of Eola1 severely affects mitochondrial translation and consequently the oxidative phosphorylation in mammalian cells. Mechanistically, EOLA1 physically interacts with mitochondrial 12S ribosomal RNA (12S mt-rRNA) and the mitochondrial Tu translation elongation factor (TUFM), facilitating efficient translation elongation of mitochondrial-encoded mRNAs. Furthermore, genetic ablation of Eola1 in mice induced severe cardiac dysfunction, demonstrating its indispensable role in maintaining heart function through mitochondrial translation control. Together, our findings establish EOLA1 as a key regulator of mitochondrial translation via its interactions with 12S mt-rRNA and TUFM, highlighting its importance in cellular metabolism and a potential role in cardiac disease pathogenesis.

molecular biology↗

Isthmin-1 is a Key Regulator of Induced Pluripotent Stem Cell-Derived Cardiomyocytes Maturation through Activation of p53 Signaling

AimsHuman induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) exhibit an immature structural and metabolic phenotype that limits their utility for cardiac disease modeling and regenerative therapy. Although multiple extrinsic strategies have been proposed to enhance iPSC-CM maturation, intrinsic molecular regulators governing this process remain incompletely defined. This study aimed to identify and characterize a novel molecular factor that promotes cardiomyocyte maturation. Methods and ResultsCross-species transcriptomic analyses comparing adult versus fetal hearts and hypertrophic cardiomyopathy versus healthy myocardium identified Isthmin-1 (ISM1) as a conserved maturation-associated gene. In human iPSC-CMs, ISM1 overexpression enhanced sarcomere organization, mitochondrial oxidative phosphorylation, ATP production, and calcium-handling properties, whereas ISM1 knockdown impaired these maturation features. RNA sequencing revealed global transcriptional reprogramming toward an adult-like state, characterized by suppression of cell-cycle-related gene programs and activation of oxidative metabolic pathways. KEGG pathway enrichment, GSVA, and GSEA consistently identified p53 signaling as the most significantly activated pathway in ISM1-overexpressing iPSC-CMs. Mechanistically, ISM1 directly interacted with p53, enhanced its protein stability, promoted nuclear localization, and increased transcription of p53 downstream targets involved in metabolic remodeling and cell-cycle exit. Pharmacological inhibition of p53 abolished ISM1-induced structural and metabolic maturation, demonstrating that ISM1 promotes cardiomyocyte maturation in a p53-dependent manner. ConclusionsISM1 is a previously unrecognized molecular driver of cardiomyocyte maturation that promotes structural, metabolic, and functional maturation of iPSC-CMs through activation of p53-dependent transcriptional programs. These findings provide new mechanistic insight into cardiomyocyte maturation and identify ISM1 as a promising target for improving the maturity of stem cell-derived cardiomyocytes. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/698535v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@15daca3org.highwire.dtl.DTLVardef@f1396dorg.highwire.dtl.DTLVardef@f13400org.highwire.dtl.DTLVardef@18d0b41_HPS_FORMAT_FIGEXP M_FIG C_FIG

developmental biology↗

Ac-FAPI-04 Promotes Anti-tumor Immunity via IL-27 Activation and Tnfaip3 Inhibition

BackgroundAccumulating evidence substantiates the capacity of targeted radionuclide therapy (TRT) to potentiate tumor immunotherapy responses. However, combining TRT and immunotherapies exhibits substantial varying efficacy, underscoring the importance of investigating mechanisms through which TRT remodels the tumor microenvironment (TME). Despite recent advancements, the specific signaling molecules mediated by TRT that regulate the functions of cytotoxic T lymphocytes (CTLs) remain unclear. MethodsGiven the demonstrated potential of -radionuclides targeting the fibroblast activation protein (FAP) in TRT, this study employed [225Ac]Ac-FAPI-04 to investigate the mechanisms of TRT-induced TME remodeling. The cytotoxic effects were first evaluated in the B16F10 cell line, followed by an assessment of its in vivo therapeutic efficacy in tumor-bearing mice. RNA-sequencing analysis was utilized to elucidate underlying signaling pathways involving immune responses. Results[225Ac]Ac-FAPI-04 significantly induced DNA double-strand breaks, further elevated ROS levels, and apoptosis in tumor cells. Transcriptomic profiling revealed that [225Ac]Ac-FAPI-04 activates the IL-27/JAK-STAT pathway to amplify interferon-mediated responses, indicating IL-27 serves as a critical regulatory signal for CTLs during TRT. A concomitant observation was the downregulation of Tnfaip3 following [225Ac]Ac-FAPI-04 treatment. Tnfaip3 is implicated in modulating CTLs behavior and intratumoral infiltration, while its downregulation was associated with the upregulation of cytotoxic effector molecules, as confirmed in this study. ConclusionThis study identifies a TRT-induced pattern of synergistic signaling mediated by IL-27 and Tnfaip3, which collectively enhances CTL recruitment and potentiates their cytotoxic function. These findings provide a detailed mechanistic rationale for strategies aimed at augmenting the therapeutic efficacy of TRT and immunotherapy combinations.

molecular biology↗

Dlx2 reprograms the transcriptome and laminar position of glia-derivedAscl1-induced interneurons

Direct lineage reprogramming of glial cells into neurons offers a promising strategy to repair diseased brain circuits, but engineering defined neuronal subtypes remains challenging. We found that a phospho-site-deficient Ascl1 variant, Ascl1SA6, but not wildtype Ascl1, induces hallmarks of parvalbumin fast-spiking interneurons, raising the question of how closely these induced neurons resemble canonical cortical interneurons and what transcriptional events underlie this process. Single-cell transcriptomic analysis revealed that Ascl1SA6-induced neurons only partially recapitulated canonical interneuron programs and failed to induce the transcription factor Dlx2 and its downstream targets. Co-expression of Dlx2 with Ascl1SA6 restored a more canonical interneuron-like transcriptome, including genes involved in migration, and resulted in neurons occupying laminar positions more typical of endogenous interneurons. These findings provide molecular insights into how Ascl1 posttranslational modifications regulate its transcriptional activity and demonstrate a strategy to engineer induced cortical interneurons that more closely resemble their native counterparts, offering a framework for layer-specific restoration of inhibitory circuits in neurological diseases.

neuroscience↗

Combined antagonism of Oncostatin M (OSM) and Interleukin 6 (IL-6) provides both anti-fibrotic and anti-inflammatory benefit in pulmonary fibrosis

Interstitial lung diseases (ILDs), including idiopathic pulmonary fibrosis (IPF) and systemic sclerosis-associated ILD (SSc-ILD), are irreversible fibrosing diseases with a mean survival time of less than 5 years for IPF. Poorly understood etiology and complex pathogenesis of these diseases have hampered the identification and development of effective therapeutics. Existing treatments can slow progressive fibrosis and lung function decline but do not stop it entirely, resulting in a minimal impact on patient survival. Thus, novel therapeutic interventions are needed. Tocilizumab, an anti-IL-6 receptor antibody, was recently approved by FDA for the treatment of SSc-ILD based on evidence demonstrating a reduction in the rate of lung function decline. In this study, we have characterized an IL-6-driven feed-forward myeloid axis contributing to lung inflammation providing a mechanistic hypothesis for tocilizumab. Concomitantly we have identified an oncostatin M (OSM) orchestrated lung injury response contributing to epithelial and endothelial cell disruption, myofibroblast activation and fibrosis. Despite dysregulated expression of IL11 in IPF, SSc-ILD, and murine models of fibrosis, we found no evidence for a pro-fibrotic role for IL-11 in vitro or in vivo. Instead, in pre-clinical models of IPF we demonstrate that antagonism of OSM alone, or to a greater degree in combination with IL-6, reduced lung fibrosis and inflammation. Translating these observations, we validated gp130:OSMR, rather than gp130:LIFR, as the dominant human receptor complex used by OSM, identifying OSMR as a potential therapeutic target to stall fibrosis.

immunology↗

A MYC Family Switch: L-MYC Drives and Maintains Neuroendocrine Lineage Programs in Prostate Cancer

Neuroendocrine prostate cancer (NEPC) is an aggressive, therapy-resistant subtype that emerges through lineage plasticity following androgen receptor (AR) pathway inhibition. Although MYC family oncogenes are central to prostate cancer progression, the role of MYCL (L-MYC) in NEPC has remained unclear. Here, we show that MYCL is selectively and robustly upregulated in NEPC patient samples and cell line models, whereas MYC is downregulated and MYCN remains low, revealing a lineage-associated MYC family switch. MYCL expression strongly correlates with the neuroendocrine lineage regulators ASCL1 and INSM1 and inversely with adenocarcinoma-associated genes. Mechanistically, MYCL activation is not driven by genomic amplification but reflects a permissive epigenetic landscape. Functionally, MYCL overexpression suppresses AR signaling and induces neuroendocrine-like transcriptional reprogramming, whereas MYCL knockdown disrupts neuroendocrine lineage identity and restores adenocarcinoma-associated gene expression, including MYC. We further identify ASCL1 and INSM1 as upstream regulators of MYCL, establishing a conserved neuroendocrine transcriptional axis. Together, these findings define MYCL as a lineage-specific regulator that drives neuroendocrine identity and plasticity in advanced prostate cancer.

cancer biology↗

A Hybridization-Based Novel DNA Storage Technology with Rapid Read Performance

The development of traditional silicon-based microelectronic storage media has increasingly lagged behind the rapidly growing demands of contemporary data storage. DNA storage, with its ultra-high information density and long-term preservation advantages, has rapidly become a research hotspot for next-generation storage technologies. However, current mainstream DNA storage methods based on synthesis and sequencing suffer from slow read/write speeds, hindering practical applications. To address this limitation, we propose an innovative hybridization-based DNA storage scheme. Through principle validation experiments, short-text read/write and error-correction tests, and high-density multi-format file storage experiments, we verify the feasibility of this approach. The proposed method exhibits high parallelism, rapid read speeds, and robust encryption capabilities, demonstrating promising application prospects in scenarios such as confidential data storage.

synthetic biology↗

Multivalent Nanobodies for Potent and Broad Neutralization of Staphylococcus aureus Toxins

Staphylococcus aureus is a leading cause of lethal bacteremia and pneumonia, which are driven by potent virulence factors such as T-cell superantigens and alpha hemolysin. S. aureus has among the highest rates of antibiotic resistance, yet no vaccines or alternative therapies are available despite decades of research. Here, we developed a repertoire of potent, high affinity nanobodies (Nbs) targeting key toxins in S. aureus infection, including superantigens (SAgs) SEB, SEC, TSST-1, and Hla. Comprehensive cryo-EM and AlphaFold3 analyses of these Nbs, which were elicited with clinical cocktail vaccines, revealed diverse neutralizing epitopes and mechanisms that provide strategic insights for immunotherapy and vaccine design. Guided by these findings, we engineered highly stable, multivalent, and multifunctional Nb constructs. These constructs included an aerosolizable trimeric Nb with enhanced neuralization activity against Hla and SEC, and an ultrapotent decameric Nb-IgG-Fc fusion construct against a wide range of major toxins in S. aureus sepsis (SEB, SEC, TSST-1, and Hla). These multifunctional Nbs demonstrated promising protective activity in murine models of pneumonia and sepsis, underscoring their potential as versatile immunotherapies that address the complex virulence profiles of S. aureus. Our work lays a foundation for precision immunotherapies beyond current treatment options to combat complex bacterial infections with multiple virulence mechanisms. Significance statementS. aureus is among the most common, antibiotic-resistant, and deadly causes of bacterial infections. We developed nanobodies against clinically significant virulence factors in S. aureus sepsis and pneumonia, including superantigens (SAgs) SEB, SEC, and TSST-1 as well as pore forming toxin Hla. These nanobodies displayed complete and potent neutralization of each toxin, exploiting a wide variety neutralizing mechanisms. Structural investigation of these diverse neutralizing nanobodies, which were elicited in llamas using clinically investigated cocktail vaccines, highlighted the importance of disrupting SAg interaction with TCR or MHCII and potential flaws in targeting poorly neutralizing conserved SAg epitopes using vaccine cocktails. Nb leads against each toxin were combined in different multivalent configurations, including an aerosolizable trimeric Nb and a half-life extended decameric Nb IgG Fc fusion construct. This work highlights multivalent nanobodies as a comprehensive yet therapeutically precise drug platform that addresses the complex virulence profiles of bacterial infectious diseases.

microbiology↗

Adipocyte-related genes analysis contributes to the prognosis of atrial fibrillation

BackgroundPatients with mild left atrial appendage fibrosis have a significantly lower rate of atrial fibrillation (AF) recurrence after 5 years compared to those with moderate or severe fibrosis, which presented a significantly increased positive area of cellular senescence. MethodsGSE135445, a RNA-sequencing dataset between epicardial adipose tissue (EAT) from patients with persistent non-valvular AF and sinus rhythm (SR), was downloaded and analyzed and hub genes were obtained. To obtain the abundance of immune cells, the CIBERSORT algorithm with 100 permutations was applied. The senescent hub genes were obtained and validated using human blood samples. ResultsA total of 164 up-regulated and 317 down-regulated DEGs was obtained between EAT from subjects with AF and SR. Using immune infiltration analysis in whole PAT related DEGs, the abundance of M0 macrophages was significantly different between PAT and SAT and 22 M0 macrophages-related DEGs were also obtained, such as MMP7, APOE, CCND1, MMP12 and APOC1. Venn diagram was utilized to obtain the same transcripts between senescence datasets and DEGs and PPI network was used to 94 overlapping genes. The top15-degree senescence adipicytes hub genes were explored. To investigate senescence-related hub genes effects on AF progression, we collected the blood samples of patients with paroxysmal atrial fibrillation and persistent atrial fibrillation. There were no significant differences at baseline characteristics and laboratory examinations at 1d after admission except heart failure history and function examinations, for instance, ALT, LDL, LVEF and LA diameters. Using univariate analysis and multivariate analysis, heart failure history, ALT and LA diameters can be prognostic factors for AF progression. BMP7, COL4A3, COL4A4, CHRD, IRX1, SPON1, ALPL, TCF19 and SIX2 were validated to be highly expressed in persistent atrial fibrillation compared to paroxysmal atrial fibrillation. ConclusionBased on our current study, BMP7, COL4A3, COL4A4, CHRD, IRX1, SPON1, ALPL, TCF19 and SIX2 were validated to be highly expressed in persistent atrial fibrillation compared to paroxysmal atrial fibrillation, which may be potential therapeutic targets to treat AF progression and improve patients prognosis.

genetics↗

Engineering chimeric DNA polymerases for DNA movable type storage

DNA-based information storage offers a promising alternative to conventional media due to its high density, long-term stability, and low energy requirements. However, its application remains hindered by synthesis costs, limited sequence length and poor scalability. DNA polymerase is a critical enzymatic tool in the DNA storage systems by enabling high-fidelity data writing and targeted sequence amplification. In this study, we engineered chimeric DNA polymerases by fusing the high-fidelity 9{degrees}N DNA polymerase with double-stranded DNA binding proteins derived from thermophilic archaea. These fusions significantly enhanced processivity, thermal stability, and salt tolerance by stabilizing enzyme-template interactions, mimicking sliding clamps while preserving catalytic efficiency. Leveraging these properties, we demonstrated precise file retrieval from a mixed oligonucleotide pool using orthogonal barcode primers. Compared with wild-type 9{degrees}N, the chimeric polymerases, particularly PLS, exhibited reduced substitution error rates and improved read accuracy. We then applied these enzymes to a DNA movable type storage system, where prefabricated DNA modules were assembled into encoding blocks. Using engineered polymerases, these blocks were recombined to enable flexible data rewriting without de novo DNA synthesis. Moreover, a multi-enzyme assembly strategy enabled the construction of kilobase-scale DNA sequences encoding a classical Chinese poem, achieving complete data recovery. All assembled fragments remained stable in E. coli over 100 generations, exhibiting the potential for in vivo storage. Collectively, our findings demonstrated the role of engineered DNA polymerases for DNA-based information storage. Moreover, this system reduced synthesis demands, supported scalable rewriting, and ensured long-term preservation, offering a practical route to sustainable, high-fidelity DNA data storage.

synthetic biology↗

Mechanisms of tumor persistence in metastatic melanoma following successful immunotherapy

Tumor dormancy is thought to enable cancer recurrence, but the settings and mechanisms of dormancy in patients are poorly characterized. Both immunogenic tumor mass dormancy, involving continued tumor cell proliferation and balanced immune-mediated cell death, and cell-level quiescence have been observed in preclinical models. Here, we demonstrate the existence of mass dormancy rather than quiescence in long-term stable residual lesions in melanoma patients treated with immune checkpoint inhibitors (ICI). In a large stable lesion subjected to detailed spatial profiling, the proportion of proliferating tumor cells is similar to that in site-matched tumors from patients progressing after ICI. Residual stable lesions from other patients, judged to contain only scar tissue upon clinical pathology review, also contained nests of dividing tumor cells surrounded by active immune cells. These findings demonstrate the presence of viable tumor cells and mass dormancy in persistent stable lesions, a finding with implications for disease monitoring and management.

cancer biology↗

PCNA Inhibition Enhances the Antitumor Activity of KRAS-Targeted Therapies in Pancreatic Cancer

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with a dismal prognosis. More than 90% of PDAC tumors harbor KRAS mutations, and several KRAS inhibitors, such as off-state, on-state, mutation-specific, and pan-RAS inhibitors, are being tested in preclinical and clinical settings. However, the response to these inhibitors as single agents is less than optimal, indicating the need to identify novel combination therapies to improve treatment outcomes. Proliferating cell nuclear antigen (PCNA) is a ring-shaped clamp protein that regulates DNA replication, repair, and resolution of transcription-replication conflict, which are critical processes for pancreatic cancer survival. AOH1996 is a first-in-class, selective PCNA inhibitor in Phase I trials. Here, we found that AOH1996 treatment is efficacious in various PDAC models in vitro. PCNA and KRAS are predicted to be synthetic lethal partners, and RNA sequencing of AOH1996-treated PDAC cells reveals enrichment of MAPK and PI3K signaling pathways. Combination of AOH1996 with KRAS inhibitors demonstrates strong synergy across KRAS G12C and G12D mutant models. Treatment with a combination of AOH1996 and KRAS inhibitors induces cell cycle arrest and apoptosis in PDAC cells. Robust antitumor activity of AOH1996 in combination with RMC-6236 was observed in PDAC tumoroids. In vivo, the combination of AOH1996 with sotorasib or MRTX1133 reduced tumor growth rates compared to single-agent therapy, with no impact on mouse body weight. Residual tumor analysis showed sustained pERK and Myc inhibition in the combination arm. In conclusion, combination of AOH1996 with KRAS inhibitors is a promising therapeutic strategy for KRAS-driven PDAC, warranting further clinical investigation.

cancer biology↗