Cardiovascular health metrics defined by Life's Essential 8 scores and subsequent macrovascular and microvascular complications in individuals with type 2 diabetes: A prospective cohort study.

AIM: To investigate the association between cardiovascular health metrics defined by Life's Essential 8 (LE8) scores and vascular complications among individuals with type 2 diabetes (T2D). MATERIALS AND METHODS: This prospective study included 11 033 participants with T2D, all devoid of macrovascular diseases (including cardiovascular and peripheral artery disease) and microvascular complications (e.g. diabetic retinopathy, neuropathy and nephropathy) at baseline from the UK Biobank. The LE8 score comprised eight metrics: smoking, body mass index, physical activity, non-high-density lipoprotein cholesterol, blood pressure, glycated haemoglobin, diet and sleep duration. Cox proportional hazards models were established to assess the associations of LE8 scores with incident macrovascular and microvascular complications. RESULTS: During a median follow-up of 12.1 years, we identified 1975 cases of incident macrovascular diseases and 1797 cases of incident microvascular complications. After adjusting for potential confounders, each 10-point increase in the LE8 score was associated with an 18% lower risk of macrovascular diseases and a 15% lower risk of microvascular complications. Comparing individuals in the highest and lowest quartiles of LE8 scores revealed hazard ratios of 0.55 (95% confidence interval 0.47-0.62) for incident macrovascular diseases, and 0.61 (95% confidence interval 0.53-0.70) for incident microvascular complications. This association remained robust across a series of sensitivity analyses and nearly all subgroups. CONCLUSION: Higher LE8 scores were associated with a lower risk of incident macrovascular and microvascular complications among individuals with T2D. These findings underscore the significance of adopting fundamental strategies to maintain optimal cardiovascular health and curtail the risk of developing diabetic vascular complications.

Upregulated LncRNA H19 Sponges MiR-106a-5p and Contributes to Aldosterone-Induced Vascular Calcification via Activating the Runx2-Dependent Pathway.

BACKGROUND: Excess aldosterone is implicated in vascular calcification (VC), but the mechanism by which aldosterone-MR (mineralocorticoid receptor) complex promotes VC is unclear. Emerging evidence indicates that long-noncoding RNA H19 (H19) plays a critical role in VC. We examined whether aldosterone-induced osteogenic differentiation of vascular smooth muscle cells (VSMCs) through H19 epigenetic modification of Runx2 (runt-related transcription factor-2) in a MR-dependent manner. METHODS: We induced in vivo rat model of chronic kidney disease using a high adenine and phosphate diet to explore the relationship among aldosterone, MR, H19, and VC. We also cultured human aortic VSMCs to explore the roles of H19 in aldosterone-MR complex-induced osteogenic differentiation and calcification of VSMCs. RESULTS: H19 and Runx2 were significantly increased in aldosterone-induced VSMC osteogenic differentiation and VC, both in vitro and in vivo, which were significantly blocked by the MR antagonist spironolactone. Mechanistically, our findings reveal that the aldosterone-activated MR bound to H19 promoter and increased its transcriptional activity, as determined by chromatin immunoprecipitation, electrophoretic mobility shift assay, and luciferase reporter assay. Silencing H19 increased microRNA-106a-5p (miR-106a-5p) expression, which subsequently inhibited aldosterone-induced Runx2 expression at the posttranscriptional level. Importantly, we observed a direct interaction between H19 and miR-106a-5p, and downregulation of miR-106a-5p efficiently reversed the suppression of Runx2 induced by H19 silencing. CONCLUSIONS: Our study clarifies a novel mechanism by which upregulation of H19 contributes to aldosterone-MR complex-promoted Runx2-dependent VSMC osteogenic differentiation and VC through sponging miR-106a-5p. These findings highlight a potential therapeutic target for aldosterone-induced VC.

Enhancing precision medicine: a nomogram for predicting platinum resistance in epithelial ovarian cancer.

BACKGROUND: This study aimed to develop a novel nomogram that can accurately estimate platinum resistance to enhance precision medicine in epithelial ovarian cancer(EOC). METHODS: EOC patients who received primary therapy at the General Hospital of Ningxia Medical University between January 31, 2019, and June 30, 2021 were included. The LASSO analysis was utilized to screen the variables which contained clinical features and platinum-resistance gene immunohistochemistry scores. A nomogram was created after the logistic regression analysis to develop the prediction model. The consistency index (C-index), calibration curve, receiver operating characteristic (ROC) curve, and decision curve analysis (DCA) were used to assess the nomogram's performance. RESULTS: The logistic regression analysis created a prediction model based on 11 factors filtered down by LASSO regression. As predictors, the immunohistochemical scores of CXLC1, CXCL2, IL6, ABCC1, LRP, BCL2, vascular tumor thrombus, ascites cancer cells, maximum tumor diameter, neoadjuvant chemotherapy, and HE4 were employed. The C-index of the nomogram was found to be 0.975. The nomogram's specificity is 95.35% and its sensitivity, with a cut-off value of 165.6, is 92.59%, as seen by the ROC curve. After the nomogram was externally validated in the test cohort, the coincidence rate was determined to be 84%, and the ROC curve indicated that the nomogram's AUC was 0.949. CONCLUSION: A nomogram containing clinical characteristics and platinum gene IHC scores was developed and validated to predict the risk of EOC platinum resistance.

Related Clinical Factors of Platinum-Based Chemotherapy Resistance in Patients with Epithelial Ovarian Cancer.

OBJECTIVE: Ovarian cancer is the second most common malignancy in women, but it is a fatal gynecological tumor. Although it has a standard treatment regimen, resistance to chemotherapy makes patients more prone to early recurrence, leading to poor survival rates. Therefore, this study investigated factors related to platinum resistance through a complete analysis of clinical data. DESIGN: Clinical data of patients with ovarian cancer were collected, and the patients were categorized into platinum-sensitive and platinum-resistant groups. By comparing the differences in clinical data between the groups, the key factors affecting platinum resistance were analyzed. PARTICIPANTS/MATERIALS, SETTING, METHODS: We collected the clinical data of patients with epithelial ovarian cancer (EOC) who were admitted to the Department of Oncology of the General Hospital of Ningxia Medical University between January 1, 2019, and December 31, 2020. We conducted univariate and multivariate analyses and evaluated overall survival and progression-free survival using the Kaplan-Meier method. RESULTS: We enrolled 161 patients with EOC, of whom 124 demonstrated platinum sensitivity and 37 demonstrated platinum resistance after the initial platinum-based chemotherapy. Univariate analyses revealed that the International Federation of Gynecology and Obstetrics (FIGO) stage, neoadjuvant chemotherapy, and Fagotti score were associated with an increased risk of platinum resistance for the first recurrence. In multivariate logistic regression analysis, only Fagotti score and neoadjuvant chemotherapy were associated with an increased risk of platinum resistance (odds ratio: 0.372 and 0.328, 95% confidence interval: 0.160-0.863 and 0.141-0.762, p = 0.021 and 0.010, respectively). LIMITATIONS: The sample size of this study was relatively small because of nonstandard treatment of some patients, the absence of clinical data, and failure of follow-up. CONCLUSIONS: Patients with EOC exhibiting platinum resistance had a very poor prognosis. The Fagotti score and neoadjuvant chemotherapy appeared to increase the risk of platinum resistance at first recurrence.

Effect of oxide interactions on chromium speciation transformation during simulated municipal solid waste incineration.

Chromium released during municipal solid waste incineration (MSWI) is toxic and carcinogenic. The removal of chromium from simulated MSWI flue gas by four sorbents (CaO, bamboo charcoal (BC), powdered activated carbon (PAC), and Al2O3) and the effects of four oxides (SiO2, Al2O3, Fe2O3, and CaO) on chromium speciation transformation were investigated. The results showed that the removal rates of total Cr by the four sorbents were Al2O3 < CaO < PAC < BC, while the removal rates of Cr(VI) by the four sorbents were Al2O3 < PAC < BC < CaO. CaO had a strong oxidizing effect on Cr(III), while BC and PAC had a better-reducing effect on Cr(VI). SiO2 was better for the reduction of Na2CrO4 and K2CrO4 above 1000°C due to its strong acidity, and the addition of CaO significantly inhibited the reduction of Cr(VI). MgCrO4 decomposed above 700°C to form MgCr2O4, and the reaction between MgCrO4 and oxides also existed in the form of a more stable trivalent spinel. Furthermore, when investigating the effect of oxides on the oxidation of Cr(III) in CrCl3, it was discovered that CaO promoted the conversion of Cr(III) to Cr(VI), while the presence of chlorine caused chromium to exist in the form of Cr(V), and increasing the content of CaO and extending the heating time facilitated the oxidation of Cr(III). In addition, silicate, aluminate, and ferrite were generated after the addition of SiO2, Al2O3, and Fe2O3, which reduced the alkalinity of CaO and had an important role in inhibiting the oxidation of Cr(III). The acidic oxides can not only promote the reduction of Cr(VI) but also have an inhibitory effect on the oxidation of Cr(III) ascribed to alkali metals/alkaline earth metals, and the proportion of acidic oxides can be increased moderately to reduce the generation of harmful substances in the hazardous solid waste heat treatment.

Two-dimensional heterostructures for photocatalytic CO2 reduction.

The photocatalysis conversion of CO2 into fuels has become an encouraging method to address climate and energy issues as a long-term solution. Single material suffers poor yield due to low light energy utilization and high recombination rate of photoinduced electron-hole pairs. It is an efficient approach to construct heterojunction through two or three materials to improve the photocatalytic performance. Recently, 2D-based heterojunction is getting popular for outstanding properties, such as special light collecting structure to enhance light harvest, intimate interface to facilitate charge transfer and separation, and large specific surface area to provide abundant reactive sites. Recently, some new 2D-based heterostructures materials (both structure and composition) have been developed with excellent performance. 2D materials exert structural and functional advantages in these fine composite photocatalysts. In this review, the literatures about the photocatalytic conversion of CO2 are mainly summarized based on overall structure, interface type and material type of 2D-based heterojunction, with special attention given to the preparation, characterization, structural advantages and reaction mechanism of novel 2D-based heterojunction. This work is in hope of offering a basis for designing improved composite photocatalyst for CO2 photoreduction.

3D Bioprinted GelMA-Nanoclay Hydrogels Induce Colorectal Cancer Stem Cells Through Activating Wnt/ß-Catenin Signaling.

Cancer stem cells (CSCs) are a rare cell population in tumors that are responsible for tumor recurrence and metastasis. They are a priority as therapeutic targets, however, assays targeting CSCs have been limited by expanding and maintaining CSCs in vitro. Here, the authors find that gelatin methacryloyl (GelMA)-nanoclay hybrid hydrogels can induce and enrich colorectal CSCs assisted by three-dimensional (3D) bioprinting. The presence of the nanoclay increases the printability, Young's modulus, pore size, and cytocompatibility of the hydrogels. Bioprinted GelMA-nanoclay hydrogels promote the formation of spheroids expressing elevated levels of the stemness markers LGR5, CD133, CD26, and SOX2. Cancer cells grown in GelMA-nanoclay hydrogel possess higher self-renewal and differentiation capacity in vitro and higher tumorigenic capacity in vivo. GelMA-nanoclay hydrogels induce CSCs by stimulating the activation of the Wnt/ß-catenin signaling pathway. Further studies demonstrate that spheroids from GelMA-nanoclay hydrogels possess increased stemness, higher consistency, yield, and sensitivity to the anti-CSC compounds compared to the classic CSC-enrichment model. Collectively, this study may provide a valuable biomaterial and method for inducing and enriching CSCs, to facilitate the effective CSC-targeting drug screening.

Reversed selectivity of photocatalytic CO2 reduction over metallic Pt and Pt(II) oxide cocatalysts.

The chemical state of Pt in cocatalysts has a major influence on the activity and selectivity of the photocatalytic reduction of CO2; however, the underlying mechanism is unclear owing to the co-existence of different Pt chemical states and mutual transformation between them. In this study, PtO/TiO2 catalysts were prepared through photodeposition and Pt/TiO2 was prepared by the photoreduction of PtO/TiO2 to avoid interference arising from co-existing Pt forms and different loading amounts. These catalysts exhibited completely reversed selectivity for CO and CH4 production during CO2 photoreduction: PtO/TiO2 tended to produce CO (100%), whereas Pt/TiO2 favored the production of CH4 (66.6%). By combining experimental analysis and theoretical calculations, the difference in selectivity was ascribed to the different charge transfer/separation and CO/H adsorption properties of PtO/TiO2 and Pt/TiO2. Photoelectric and photoluminescence (PL) analysis showed that Pt was more advantageous to the photogenerated carrier separation compared with PtO, which was conducive to the multi-electron CH4 reduction reaction. Fourier transform-infrared spectroscopy, temperature-programmed desorption/temperature-programmed reduction, and density functional theory calculations indicated that the adsorption of CO and hydrogen on Pt was stronger than that on PtO, which favored the further reduction of CO to CH4. Based on the above results, a mechanism was proposed to explain the reversed selectivity of the photocatalytic reduction of CO2 over Pt/TiO2 and PtO/TiO2.

The molecular architecture of photoreceptor phosphodiesterase 6 (PDE6) with activated G protein elucidates the mechanism of visual excitation.

Photoreceptor phosphodiesterase 6 (PDE6) is the central effector of the visual excitation pathway in both rod and cone photoreceptors, and PDE6 mutations that alter PDE6 structure or regulation can result in several human retinal diseases. The rod PDE6 holoenzyme consists of two catalytic subunits (Pαß) whose activity is suppressed in the dark by binding of two inhibitory γ-subunits (Pγ). Upon photoactivation of rhodopsin, the heterotrimeric G protein (transducin) is activated, resulting in binding of the activated transducin α-subunit (Gtα) to PDE6, displacement of Pγ from the PDE6 active site, and enzyme activation. Although the biochemistry of this pathway is understood, a lack of detailed structural information about the PDE6 activation mechanism hampers efforts to develop therapeutic interventions for managing PDE6-associated retinal diseases. To address this gap, here we used a cross-linking MS-based approach to create a model of the entire interaction surface of Pγ with the regulatory and catalytic domains of Pαß in its nonactivated state. Following reconstitution of PDE6 and activated Gtα with liposomes and identification of cross-links between Gtα and PDE6 subunits, we determined that the PDE6-Gtα protein complex consists of two Gtα-binding sites per holoenzyme. Each Gtα interacts with the catalytic domains of both catalytic subunits and induces major changes in the interaction sites of the Pγ subunit with the catalytic subunits. These results provide the first structural model for the activated state of the transducin-PDE6 complex during visual excitation, enhancing our understanding of the molecular etiology of inherited retinal diseases.

Selective-area growth of periodic nanopyramid light-emitting diode arrays on GaN/sapphire templates patterned by multiple-exposure colloidal lithography.

Gallium nitride-based nanopyramid light-emitting diodes are a promising technology to achieve highly efficient solid-state lighting and beyond. Here, periodic nanopyramid light-emitting diode arrays on gallium nitride/sapphire templates were fabricated by selective-area metalorganic chemical vapor deposition and multiple-exposure colloidal lithography. The electric field intensity distribution of incident light going through polystyrene microspheres and photoresist are simulated using finite-different time-domain method. Nitrogen as the carrier gas and a low V/III ratio (ratio of molar flow rate of group-V to group-III sources) are found to be important in order to form gallium nitride nanopyramid. In addition, a broad yellow emission in photoluminescence and cathodoluminescence spectra were observed. This phenomena showed the potential of nanopyramid light-emitting diodes to realize long wavelength visible emissions.

Multiple-exposure colloidal lithography for enhancing light output of GaN-based light-emitting diodes by patterning Ni/Au electrodes.

Ni/Au electrodes with single, twined and triplet hole array patterns light-emitting diodes have been fabricated by multiple-exposure colloidal lithography. It is found that 45.6%, 83.6% and 15.5% improvement in light output at 350 mA has been achieved by patterning Ni/Au electrodes with single, twined, triplet hole arrays. In addition, patterned Ni/Au LEDs possess much larger view angles than non-patterned ones due to scattering effects of light around the holes, especially for triplet hole array patterned Ni/Au LEDs. Our proposed method for fabricating multiple holes structure would be very promising to improve light output power of LEDs when using advanced electrodes.

Synthesis, characterization and enhanced photocatalytic CO2 reduction activity of graphene supported TiO2 nanocrystals with coexposed {001} and {101} facets.

It is known that the combination of TiO2 and graphene and the control of TiO2 crystal facets are both effective routes to improve the photocatalytic performance of TiO2. Here, we report the synthesis and the photocatalytic CO2 reduction performance of graphene supported TiO2 nanocrystals with coexposed {001} and {101} facets (G/TiO2-001/101). The combination of TiO2 and graphene enhanced the crystallinity of TiO2 single nanocrystals and obviously improved their dispersion on graphene. The "surface heterojunction" formed by the coexposed {001} and {101} facets can promote the spatial separation of photogenerated electrons and holes toward different facets and the supports of graphene can further enhance the separation through accelerated electron migration from TiO2 to graphene. The G/TiO2-001/101 exhibited high photocatalytic CO2-reduction activity with a maximum CO yield reaching 70.8 µmol g(-1) h(-1). The enhanced photocatalytic activity of the composites can be attributed to their high surface area, good dispersion of TiO2 nanoparticles, and effective separation of excited charges due to the synergy of graphene supports and the co-exposure of {001} and {101} facets.

Layer-by-Layer Approach to (2+1)D Photonic Crystal Superlattice with Enhanced Crystalline Integrity.

Large-area polystyrene (PS) colloidal monolayers with high mechanical strength are created by a combination of the air/water interface self-assembly and the solvent vapor annealing technique. Layer-by-layer (LBL) stacking of these colloidal monolayers leads to the formation of (2+1)D photonic crystal superlattice with enhanced crystalline integrity. By manipulating the diameter of PS spheres and the repetition period of the colloidal monolayers, flexible control in structure and stop band position of the (2+1)D photonic crystal superlattice has been realized, which may afford new opportunities for engineering photonic bandgap materials. Furthermore, an enhancement of 97.3% on light output power of a GaN-based light emitting diode is demonstrated when such a (2+1)D photonic crystal superlattice employed as a back reflector. The performance enhancement is attributed to the photonic bandgap enhancement and good angle-independence of the (2+1)D photonic crystal superlattice.

Super-aligned carbon nanotubes patterned sapphire substrate to improve quantum efficiency of InGaN/GaN light-emitting diodes.

In this paper, the high performance GaN-based light-emitting diodes (LEDs) on carbon-nanotube-patterned sapphire substrate (CNPSS) by metal-organic chemical vapor deposition (MOCVD) are demonstrated. By studying the mechanism of nucleation, we analyze the reasons of the crystal quality improvement induced by carbon nanotubes (CNTs) in different growth process. Combining with low temperatures photoluminescence (PL) measurements and two-dimensional (2D) finite difference time-domain (FDTD) simulation results, we conclude that the improvement of optical properties and electrical properties of CNPSS mainly originates from the improvement of the internal quantum efficiency (IQE) due to decreased dislocation density during nano-epitaxial growth on CNPSS. Additionally, in order to reduce the light absorption characteristics of CNTs, different time annealing under the oxygen environment is carried out to remove part of CNTs. Under 350 mA current injections, the light output power (LOP) of CNPSS-LED annealed 2 h and 10 h exhibit 11% and 6% enhancement, respectively, compared to that of the CNPSS-LED without annealing. Therefore, high temperature annealing can effectively remove parts of CNTs and further increase the LOP, while overlong annealing time has caused degradation of the quantum well resulting in the attenuation of optical power.

Enhanced performance of InGaN-based light emitting diodes through a special etch and regrown process in n-GaN layer.

We reported that the peak efficiency together with the efficiency droop in InGaN-based light emitting diodes could be effectively modified through a simple and low-cost etch-regrown process in n-GaN layer. The etched n-GaN template contained pyramid arrays with inclined side planes. The following lateral overgrowth process from the etched n-GaN template substantially reduced the edge dislocation density and residential compressive strain in epilayers. The efficiency droop of LED samples thus could be modified due to the reduced polarization field, resulting from the strain relaxation in epilayers. What is more, the peak efficiency and reverse current leakage were also modified due to the reduction of dislocations.

Towards the Clinical Translation of 3D PLGA/ß-TCP/Mg Composite Scaffold for Cranial Bone Regeneration.

Recent years have witnessed the rapid development of 3D porous scaffolds with excellent biocompatibility, tunable porosity, and pore interconnectivity, sufficient mechanical strength, controlled biodegradability, and favorable osteogenesis for improved results in cranioplasty. However, clinical translation of these scaffolds has lagged far behind, mainly because of the absence of a series of biological evaluations. Herein, we designed and fabricated a composite 3D porous scaffold composed of poly (lactic-co-glycolic) acid (PLGA), ß-tricalcium phosphate (ß-TCP), and Mg using the low-temperature deposition manufacturing (LDM) technique. The LDM-engineered scaffolds possessed highly porous and interconnected microstructures with a porosity of 63%. Meanwhile, the scaffolds exhibited mechanical properties close to that of cancellous bone, as confirmed by the compression tests. It was also found that the original composition of scaffolds could be maintained throughout the fabrication process. Particularly, two important biologic evaluations designed for non-active medical devices, i.e., local effects after implantation and subchronic systemic toxicity tests, were conducted to evaluate the local and systemic toxicity of the scaffolds. Additionally, the scaffolds exhibited significant higher mRNA levels of osteogenic genes compared to control scaffolds, as confirmed by an in vitro osteogenic differentiation test of MC3T3-E1 cells. Finally, we demonstrated the improved cranial bone regeneration performance of the scaffolds in a rabbit model. We envision that our investigation could pave the way for translating the LDM-engineered composite scaffolds into clinical products for cranial bone regeneration.

[3D tumor spheroids promote activation, expansion, and anti-tumor effects of tumor-infiltrating lymphocytes in vitro].

The adoptive immunotherapy mediated by tumor-infiltrating lymphocytes (TILs) has shown definite efficacy against various solid tumors. However, the inefficiency of the conventional method based on in vitro expansion of TILs fails to achieve the cell count and high tumor-killing activity required for therapeutic purposes. This study investigated the effect of 3D tumor spheroids on the activation and expansion of TILs in vitro, aiming to provide a novel approach for the expansion of TILs. We procured TILs and primary tumor cells from surgical samples of lung cancer patients and then compared the impacts of lung cancer cell line NCI-H1975 and primary lung cancer cells cultured under 2D and 3D conditions on the activation, expansion, and anti-tumor activity of TILs. Furthermore, we added the programmed cell death protein 1 (PD-1) antibody into the co-culture of primary tumor cells and TILs within a 3D environment to assess the effects of the antibody on TILs. The results showed that compared with 2D cultured tumor cells, the 3D cultured H1975 cells significantly enhanced the expansion of TILs, increasing the proportion of CD3+/CD8+ cells in TILs to 61.6%. The 3D primary tumor model also enhanced the proportion of CD3+/CD8+ cells in TILs (45.5%, 54.4%), induced apoptosis of tumor epithelial cells and decreased the overall tumor cells survival rate (16.7%) after co-culture. PD-1 antibodies further improved the in vitro expansion capacity of TILs mediated by 3D tumor spheroids, resulting in the proportions of 50.9% and 57.0% for CD3+/CD8+ cells and enhancing the antitumor activity significantly (reducing the overall tumor survival rate to 9.36%). In summary, the use of 3D tumor spheroids significantly promoted the expansion and improved the anti-tumor effect of TILs, and the use of the PD-1 antibody further promoted the expansion and tumor-killing effect of TILs.

Gastric Cancer Assembloids Derived from Patient-Derived Xenografts: A Preclinical Model for Therapeutic Drug Screening.

The construction of reliable preclinical models is crucial for understanding the molecular mechanisms involved in gastric cancer and for advancing precision medicine. Currently, existing in vitro tumor models often do not accurately replicate the human gastric cancer environment and are unsuitable for high-throughput therapeutic drug screening. In this study, droplet microfluidic technology is employed to create novel gastric cancer assembloids by encapsulating patient-derived xenograft gastric cancer cells and patient stromal cells in Gelatin methacryloyl (GelMA)-Gelatin-Matrigel microgels. The usage of GelMA-Gelatin-Matrigel composite hydrogel effectively alleviated cell aggregation and sedimentation during the assembly process, allowing for the handling of large volumes of cell-laden hydrogel and the uniform generation of assembloids in a high-throughput manner. Notably, the patient-derived xenograft assembloids exhibited high consistency with primary tumors at both transcriptomic and histological levels, and can be efficiently scaled up for preclinical drug screening efforts. Furthermore, the drug screening results clearly demonstrated that the in vitro assembloid model closely mirrored in vivo drug responses. Thus, these findings suggest that gastric cancer assembloids, which effectively replicate the in vivo tumor microenvironment, show promise for enabling more precise high-throughput drug screening and predicting the clinical outcomes of various drugs.

A patient-specific lung cancer assembloid model with heterogeneous tumor microenvironments.

Cancer models play critical roles in basic cancer research and precision medicine. However, current in vitro cancer models are limited by their inability to mimic the three-dimensional architecture and heterogeneous tumor microenvironments (TME) of in vivo tumors. Here, we develop an innovative patient-specific lung cancer assembloid (LCA) model by using droplet microfluidic technology based on a microinjection strategy. This method enables precise manipulation of clinical microsamples and rapid generation of LCAs with good intra-batch consistency in size and cell composition by evenly encapsulating patient tumor-derived TME cells and lung cancer organoids inside microgels. LCAs recapitulate the inter- and intratumoral heterogeneity, TME cellular diversity, and genomic and transcriptomic landscape of their parental tumors. LCA model could reconstruct the functional heterogeneity of cancer-associated fibroblasts and reflect the influence of TME on drug responses compared to cancer organoids. Notably, LCAs accurately replicate the clinical outcomes of patients, suggesting the potential of the LCA model to predict personalized treatments. Collectively, our studies provide a valuable method for precisely fabricating cancer assembloids and a promising LCA model for cancer research and personalized medicine.

Functional mapping of interacting regions of the photoreceptor phosphodiesterase (PDE6) γ-subunit with PDE6 catalytic dimer, transducin, and regulator of G-protein signaling9-1 (RGS9-1).

The cGMP phosphodiesterase (PDE6) involved in visual transduction in photoreceptor cells contains two inhibitory γ-subunits (Pγ) which bind to the catalytic core (Pαß) to inhibit catalysis and stimulate cGMP binding to the GAF domains of Pαß. During visual excitation, interaction of activated transducin with Pγ relieves inhibition. Pγ also participates in a complex with RGS9-1 and other proteins to accelerate the GTPase activity of activated transducin. We studied the structural determinants for these important functions of Pγ. First, we identified two important sites in the middle region of Pγ (amino acids 27-38 and 52-54) that significantly stabilize the overall binding affinity of Pγ with Pαß. The ability of Pγ to stimulate noncatalytic cGMP binding to the GAF domains of PDE6 has been localized to amino acids 27-30 of Pγ. Transducin activation of PDE6 catalysis critically depends on the presence of Ile54 in the glycine-rich region of Pγ in order to relieve inhibition of catalysis. The central glycine-rich region of Pγ is also required for transducin to increase cGMP exchange at the GAF domains. Finally, Thr-65 and/or Val-66 of Pγ are critical residues for Pγ to stimulate GTPase activity of transducin in a complex with RGS9-1. We propose that the glycine-rich region of Pγ is a primary docking site for PDE6-interacting proteins involved in the activation/inactivation pathways of visual transduction. This functional mapping of Pγ with its binding partners demonstrates the remarkable versatility of this multifunctional protein and its central role in regulating the activation and lifetime of visual transduction.