Category Archives: 研究發表

趙瑞益教授研究團隊發表研究成果於Adv Healthc Mater.

連結網址：https://pubmed.ncbi.nlm.nih.gov/42266017/

Abstract

Targeted protein degradation has emerged as a transformative therapeutic modality in cancer therapy, enabling the elimination of undruggable proteins and overcoming conventional small-molecule resistance. Therapeutic approaches such as endocytosis-targeting chimera (ENDOTAC) and autophagy-targeting chimera (AUTOTAC) facilitate undruggable protein elimination. Here, functionalized fluorescent nanodiamond-targeting chimera (FND-TAC) reveals therapeutic protein clearance and tumor suppression through ENDOTAC linked to AUTOTAC networks. SQSTM1 is a key autophagy receptor mediating ENDOTAC linked to AUTOTAC for targeting protein degradation and cancer therapy. FND is a nanocarrier with stable fluorescence, enabling real-time tracking of protein degradation and tumorigenesis. Pull-down proteomics and STRING network analyses identified SQSTM1 as a central mediator linking Rab5/Rab7-mediated FND endocytosis through ENDOTAC to AUTOTAC. Rab5/Rab7 ubiquitination, coordinated activation of the SQSTM1 PB1, LIR, and UBA domains, and SQSTM1 S403 phosphorylation through AUTOTAC enabled FND nanoparticulosome formation and subsequent nanoparticulophagy. Moreover, SQSTM1-mediated LAMP1/2 organization linked AUTOTAC to LYTAC for lysosomal targeting. Functionalization with the therapeutic PD-L1 monoclonal antibody atezolizumab (ATZ) to form FND-ATZ demonstrated that ATZ-targeted PD-L1 was delivered to lysosomes for clearance and cancer cell death. Furthermore, FND-ATZ highlighted therapeutic PD-L1 clearance and tumor suppression through ENDOTAC linked to AUTOTAC networks. Real-time observation of FND-TAC reveals that ENDOTAC-AUTOTAC networks mediate therapeutic protein clearance and tumor inhibition.

趙啟宏副教授研究團隊發表研究成果於Cell Death Dis.

連結網址：https://pubmed.ncbi.nlm.nih.gov/42436149/

Abstract

Enolase 2 (ENO2) is a neuron-specific glycolytic enzyme whose expression is elevated in aggressive breast cancers, yet its enzymatic and biological contributions to triple-negative breast cancer (TNBC) progression remain incompletely defined. Here, we demonstrate that ENO2 sustains cancer stem cell (CSC) properties and metastatic competence through a phosphoenolpyruvate (PEP)-dependent metabolic axis. Elevated ENO2 expression correlated with advanced tumor grade and poor clinical outcomes in TNBC cohorts, underscoring its clinical relevance. Genetic depletion of ENO2 impaired aerobic glycolysis and oxidative phosphorylation, reduced migration, invasion, and CSC frequency, and suppressed tumor growth and pulmonary metastasis in orthotopic models. Mechanistically, exogenous PEP or pyruvate restored CSC-associated traits and invasiveness in ENO2-deficient cells, supporting the functional involvement of ENO2-derived metabolites in CSC maintenance. Reconstitution with wild-type ENO2, but not a catalytically impaired mutant, restored CSC properties, invasiveness, and metastatic colonization, establishing that ENO2 catalytic activity is required for these malignant traits. Further analysis revealed that PKM2 perturbation preferentially attenuated PEP-mediated rescue while largely sparing pyruvate-mediated rescue, supporting a functional PEP-PKM2-pyruvate axis in CSC regulation. Consistently, PKM2 depletion partially blunted ENO2-mediated rescue; however, residual rescue despite PKM2 perturbation suggested additional PKM2-independent PEP-responsive mechanisms. Importantly, pharmacological inhibition of enolase with POMHEX phenocopied genetic ENO2 loss and suppressed CSC maintenance in vitro and tumor growth in vivo. Taken together, these findings identify the ENO2-driven PEP-dependent metabolic axis as a mechanistic link between metabolic reprogramming, cancer stemness, and metastasis, revealing a therapeutically actionable metabolic vulnerability in TNBC.

吳東昆教授、張晉源副教授研究團隊發表研究成果於J Am Chem Soc.

連結網址：https://pubmed.ncbi.nlm.nih.gov/41779877/

Abstract

The biosynthesis of α-cyclopiazonic acid (α-CPA) is notable for generating a complex pentacyclic scaffold using a minimal three-enzyme pathway. The final step, catalyzed by CpaO, converts linear β-CPA into α-CPA through an enigmatic oxidative cyclization. Here, we report the structural and mechanistic characterization of CpaO. X-ray crystallography reveals a three-domain architecture belonging to the flavin-dependent amine oxidase (FAO) superfamily, while CpaO represents a previously undescribed subfamily distinguished by an essential, covalently linked FAD (8α-N¹-histidyl) and divergent substrate-binding domains. High-resolution structures of the CpaO/β-CPA complex, validated by mutagenesis, identify key active-site residues (His165, Trp317, Asp412, Tyr283) that anchor the substrate. Combined structural, mutational, and molecular dynamics analyses further suggest distinct yet cooperative roles for Tyr283 and Ser167 in modulating substrate access and subsequent binding. Derived from these data, we propose a stereospecific mechanism initiated by FAD-mediated hydride abstraction, which triggers a bicyclization cascade to form the final C and D rings. This study resolves a long-standing biosynthetic mystery and expands the catalytic repertoire of flavoenzymes, offering a template for the chemoenzymatic synthesis of complex indole alkaloids.

李明家副教授研究團隊發表研究成果於Angewandte Chemie International Edition

連結網址：https://onlinelibrary.wiley.com/doi/10.1002/anie.4704601

Abstract
Circularly polarized luminescence (CPL) has emerged as a key optical property with chiral-photonic and optoelectronic applications. However, conventional CPL systems rely on synthetically demanding chiral luminophores that emit at fixed wavelengths, which limits tunability and efficiency. In this study, we demonstrate that achiral or racemic luminophores co-encapsulated with photosensitizers in the helical nanocavities of chiral silica exhibit upconversion CPL (UC-CPL). The chiral silica, which is derived from polymethylvinylsiloxane comprising polyhedral oligomeric silsesquioxane decorated with enantiomeric N-(tert-butoxycarbonyl)cysteine methyl ester moieties (PMVS-POSS-Cys), has a helical structure with a preferred-handedness that facilitates efficient triplet–triplet energy transfer (TTET) and triplet–triplet annihilation (TTA). The resulting materials exhibit distinct CPL and UC-CPL signals when excited at 365 and 532 nm, respectively, despite the achiral nature of the luminophore. This study unprecedentedly demonstrates UC-CPL from achiral emitters within chiral silica matrices, which is achieved through chiral induction in the solid-state helical silica frameworks. The strategy described herein provides a general and versatile platform for developing energy-efficient, wavelength-tunable chiral-photonic materials without the need for elaborate chiral syntheses.

李明家副教授研究團隊發表研究成果於ACS Applied Engineering Materials

連結網址：https://pubs.acs.org/doi/full/10.1021/acsaenm.6c00136

Abstract

In this study, the crystallization behavior of poly(ε-caprolactone) (PCL) under MEW processing conditions was systematically investigated to clarify its role in governing fiber morphology and functional properties. By systematically varying critical processing parameters, both the degree of crystallinity and the orientation of crystalline domains were modulated and quantitatively assessed using polarized light microscopy (PLM), scanning electron microscopy (SEM), wide-angle X-ray diffraction (WAXD), and small-angle X-ray scattering (SAXS). The findings reveal that crystallization kinetics are strongly dictated by the thermal and shear histories imposed during MEW, resulting in pronounced differences in surface topography, interfiber fusion, and layer-to-layer stacking. These microstructural transformations are further correlated with changes in alter surface roughness and wetting behavior, indicating that controlled crystallization can be exploited as a key process-driven design parameter linking fiber microstructure to interfacial and mechanical performance in MEW-fabricated constructs.

蕭育源教授及朱智瑋研究團隊發表研究成果於Nucleic Acids Res.

連結網址：https://pubmed.ncbi.nlm.nih.gov/41533589/

Abstract

In nucleic acid-binding proteins, short linear motifs (SLiMs)-an important subclass of intrinsically disordered regions (IDRs)-offer diverse opportunities for therapeutic intervention, yet their structural and functional roles remain largely elusive. Away from the active site of cancer immunotherapy target exonuclease TREX1, a novel modulation site formed by the intrinsically disordered α7-α8 loop is discovered by X-ray crystallography with newly identified inhibitors. Despite that the structure of α7-α8 loop upon binding-coupled disordered-to-ordered transition is inhibitor specific, a pattern of multivertex clamping is consistently observed. Mechanistically, the fuzzy TREX1-inhibitor interactions elucidated by structural analysis and molecular dynamics simulations reveal an ensemble of chemical-scale amphiphilic units for inhibitor moieties to anchor to. Functional assays confirm that our newly identified inhibitors disrupt the DNA binding and immunosuppressive activity of TREX1, establishing α7-α8 loop as a druggable SLiM. This work provides a first collection of atomic details for small-molecule inhibition involving a DNA-binding SLiM, and the mechanistic principles uncovered here may be generalized to targeting IDRs in cancer immunotherapy.

趙瑞益教授研究團隊發表研究成果於J Adv Res.

連結網址：https://pubmed.ncbi.nlm.nih.gov/41730411/

Abstract

Introduction: Biomolecular condensates are membraneless organelles functionally involved in diverse processes, including cancer progression. Sequestosome-1 (SQSTM1)/p62 regulates liquid-liquid phase separation to drive biomolecular condensates, while programmed death-ligand 1 (PD-L1) promotes cancer cell growth. Although SQSTM1 has been shown to form condensates with several complexes, its full range of partners remains incompletely characterized.

Objectives: This study aimed to investigate the role of SQSTM1 in mediating PD-L1 condensate formation and its contribution to lung tumorigenesis.

Methods: The SQSTM1-PD-L1 interaction and condensate colocalization were examined by immunoprecipitation and immunofluorescence. Truncated SQSTM1 constructs validate the interacting domain with PD-L1, which is supported by computational AlphaFold3 analysis. We applied 1,6-hexanediol, FDA-approved PD-L1 antibody Atezolizumab, and CRISPR/Cas9-based SQSTM1 knockout to disrupt SQSTM1/PD-L1 biomolecular condensates, along with in vitro and in vivo experiments. The Survivin levels were analyzed using real-time quantitative PCR and immunoblotting. Clinical correlations of SQSTM1 and PD-L1 expression with prognosis were assessed using public datasets.

Results: Our research demonstrate that SQSTM1, by its Phox1 and Bem1 (PB1) domain, directly interacts with PD-L1 to facilitate the SQSTM1/PD-L1 biomolecular condensate formation. The formation of SQSTM1/PD-L1 condensates prevents PD-L1 from ubiquitin-proteasome-mediated degradation and stabilizes PD-L1 levels. Deletion of the PB1 domain in SQSTM1 inhibits the formation of PD-L1 biomolecular condensates and induces PD-L1 K48 ubiquitination for protein degradation. Disruption of SQSTM1/PD-L1 condensates with 1,6-hexanediol induces PD-L1 proteolysis and reduces cell viability. Knockout of SQSTM1 disrupts PD-L1 condensate formation, downregulates PD-L1 protein levels, and attenuates tumor growth. Treatment with PD-L1 antibody drug Atezolizumab inhibits SQSTM1/PD-L1 condensates and suppresses tumor formation. Clinically, high SQSTM1 and PD-L1 expression correlated with poor prognosis in lung cancer patients.

Conclusion: Together, the SQSTM1 directly interacts with non-membrane-associated PD-L1 to drive the formation of SQSTM1/PD-L1 biomolecular condensates, which prevent PD-L1 degradation, promote lung tumorigenesis, and serve as a potential therapeutic target in lung cancers.

Keywords: Biomolecular condensate; PB1 domain; PD-L1; SQSTM1; Tumorigenesis.

陳亭妏副教授研究團隊發表研究成果於J Hepatol.

連結網址：https://pubmed.ncbi.nlm.nih.gov/39763896/

Abstract

Most malignant hepatocellular tumors in children are classified as either hepatoblastoma (HB) or hepatocellular carcinoma (HCC), but some tumors demonstrate features of both HB and HCC^1-3. These tumors have been recognized under a provisional diagnostic category by the World Health Organization and are distinguished from HB and HCC by a combination of histological, immunohistochemical, and molecular features^4-6. Their outcomes and cellular composition remain an open question^7-9. The heterogeneous histological and molecular profiles of hepatoblastomas with carcinoma features (HBCs)⁴ may result from cells with combined HB and HCC characteristics (HBC cells) or from mixtures of cells displaying either HB or HCC signatures. We used multiomics profiling to show that HBCs are mixtures of HB, HBC, and HCC cell types. HBC cells are more chemoresistant than HB cells, and their chemoresistance-a driver of poor outcomes^10-12-is determined by their cell types, genetic alterations, and embryonic differentiation stages. We showed that the prognosis of HBCs is significantly worse than that of HBs. We also showed that HBC cells are derived from HB cells at early hepatoblast differentiation stages, that aberrant activation of WNT-signaling initiates HBC transformation, and that WNT inhibition promotes differentiation and increases sensitivity to chemotherapy. Furthermore, our analysis revealed that each HBC is the product of multiple HB-to-HBC and HBC-to-HCC transitions. Thus, multiomics profiling of HBCs provided key insights into their biology and resolved major questions regarding the etiology of these childhood liver tumors.

張晉源副教授研究團隊發表研究成果於J Am Chem Soc.

連結網址：https://pubmed.ncbi.nlm.nih.gov/41779877/

Abstract

The biosynthesis of α-cyclopiazonic acid (α-CPA) is notable for generating a complex pentacyclic scaffold using a minimal three-enzyme pathway. The final step, catalyzed by CpaO, converts linear β-CPA into α-CPA through an enigmatic oxidative cyclization. Here, we report the structural and mechanistic characterization of CpaO. X-ray crystallography reveals a three-domain architecture belonging to the flavin-dependent amine oxidase (FAO) superfamily, while CpaO represents a previously undescribed subfamily distinguished by an essential, covalently linked FAD (8α-N¹-histidyl) and divergent substrate-binding domains. High-resolution structures of the CpaO/β-CPA complex, validated by mutagenesis, identify key active-site residues (His165, Trp317, Asp412, Tyr283) that anchor the substrate. Combined structural, mutational, and molecular dynamics analyses further suggest distinct yet cooperative roles for Tyr283 and Ser167 in modulating substrate access and subsequent binding. Derived from these data, we propose a stereospecific mechanism initiated by FAD-mediated hydride abstraction, which triggers a bicyclization cascade to form the final C and D rings. This study resolves a long-standing biosynthetic mystery and expands the catalytic repertoire of flavoenzymes, offering a template for the chemoenzymatic synthesis of complex indole alkaloids.

高雅婷副教授研究團隊發表研究成果於Phys Chem Chem Phys.

連結網址：https://pubmed.ncbi.nlm.nih.gov/41431324/

Abstract

Fatty acid photodecarboxylases (FAPs) utilize blue light to convert fatty acids into Cn-1 hydrocarbons and CO2, providing a sustainable route to photobiocatalytic fuel production. Despite increasing interest, substrate-dependent phenomena-such as binding affinity, initial electron-transfer (ET) dynamics, and the influence of residues near the FAD cofactor-remain insufficiently characterized. Here, we systematically analyzed substrate binding, substrate-to-product conversion, and initial ET dynamics for three fatty acids of varying chain lengths using two cosolvent buffer systems. Ethanol improved substrate solubility but increased enzyme flexibility, leading to an extended FAD-substrate distance (DFAD-substrate) and diminished fluorescence quenching. Fluorescence titrations revealed chain-length-dependent binding, with palmitic acid (C16) and arachidic acid (C20) exhibiting stronger binding than lauric acid (C12). A fluorometric assay enabled quantification of product formation and catalytic half-lives, revealing that the fastest turnover arises from binding competition between substrate and product. Time-resolved fluorescence measurements further demonstrate that ET rates increase with substrate chain length. Longer fatty acids position their carboxylate groups closer to the FAD cofactor, reducing DFAD-substrate and accelerating the initial ET step. Finally, targeted mutagenesis at residue N170-located adjacent to the isoalloxazine ring-shows that local active-site interactions strongly modulate the partitioning of the excited 1FAD* pathways, thereby tuning photocatalytic efficiency and photostability.