Statin therapy and upper tract urothelial carcinoma risk in hyperlipidemic patients with chronic kidney disease and end-stage kidney disease, a population-based 17-year follow-up study.

BACKGROUND: The upper tract urothelial carcinoma (UTUC) risk associated with statin therapy in hyperlipidemic patients with chronic kidney disease (CKD) and end-stage kidney disease (ESKD) remains obscure. AIM: This retrospective cohort study investigated the UTUC risk for hyperlipidemic patients with CKD or ESKD associated with statin therapy. METHODS: From the national insurance claims data of Taiwan, we identified hyperlipidemic patients and established three pairs of statin users and non-users sub-cohorts matched by propensity scores: 401,490 pairs with normal kidney function, 37,734 pairs with CKD, and 6271 pairs with ESKD. Incidence rates and hazard ratio (HR) of UTUC were estimated, by the end of 2016, between statin and non-statin cohorts, and between hydrophilic statins users and lipophilic statins users. Time-dependent model estimated adjusted HR, and sub-distribution HR (sHR) accounting for the competing risk of deaths. RESULTS: The statin-users with ESKD were at increased UTUC risk (sHR 1.98; 95% confidence interval (CI), 1.28-3.06), significant for younger patients (40-64 years). The incidence was twofold greater in women than in men (31.8 versus 15.9 per 10,000 person-years). Receiving lipophilic statins was associated with increased UTUC risk in CKD and ESKD patients, while receiving hydrophilic statins was associated with increased UTUC risk in ESKD patients. CONCLUSIONS: Patients with ESKD receiving statin were at an increased UTUC risk, significant for younger group (<65 y/o). The positive associations between UTUC and statin persisted in both genders with ESKD, and in therapy with either lipophilic statins or hydrophilic statins. Statin users with ESKD deserve attention for UTUC prevention.

Influence of Hospice Palliative Care on Medical Service Usage in Prolonged Mechanical Ventilation Cases: A Nationwide Cohort Study.

BACKGROUND: Numerous studies have demonstrated that hospice palliative care interventions for cancer patients can reduce health care utilzation. In Taiwan, 20-25% of patients who require mechanical ventilation are using prolonged mechanical ventilation (PMV); however, only a limited number of studies have addressed the effectiveness of hospice palliative care for these patients. This study investigated the impact of hospice palliative care utilization on medical utilization among subjects using PMV. METHODS: By using the health insurance database of a nationwide population-based study, we identified subjects who had been on mechanical ventilation for > 21 d, were age ≥18 y between 2009 and 2017, and had received hospice palliative care. The control group was formed through 1:1 matching by using propensity scoring after excluding patients who had participated in palliative care for <15 d or for >181 d. Furthermore, we used a conditional logistic regression analysis to investigate the incidence of ICU admission, emergency department presentation, and cardiopulmonary resuscitation within 14 d before death. RESULTS: A total of 186,533 new subjects receiving PMV age ≥ 18 y were admitted between 2009 and 2017. In addition, the number of subjects receiving palliative care increased annually, rising from 0.6% in 2009 to 41.33% in 2017. The emergency department visits (odds ratio [OR] 0.68, 95% CI 0.63-0.74), ICU admission (OR 0.59, 95% CI 0.53-0.66), cardiopulmonary resuscitation (OR 0.40, 95% CI 0.35-0.46), and total hospitalization cost ($1,319.91 ± $1,821.66 versus $1,544.37 ± $2,309.27 [$USD], P < .001) were significant lower in the palliative care group. CONCLUSIONS: Subjects undergoing PMV while receiving hospice palliative care experienced significant reductions in total hospitalization costs, ICU admissions, cardiopulmonary resuscitation, and medical expenses within 14 d before death.

Proportion trends, cancer stage, and survival of patients with cancer diagnosed through emergency and nonemergency departments: a nationwide cohort study.

Introduction: To reduce mortality, the Taiwan government has vigorously promoted free cancer screening and preventive health screening services. Cancers are usually advanced by the time they are discovered in the emergency department. Through this study, we aimed to understand the characteristics of cancer patients diagnosed through the emergency department and thus identify high-risk populations by comparing cancer staging and survival rates in patients diagnosed in the emergency department and those diagnosed in the non-emergency department. Methods: The retrospective study enrolled a total of 389,043 patients over the age of 20 who were newly diagnosed with one of the five major cancers (including lung cancer, colorectal cancer, breast cancer, prostate cancer, and oral cancer) between 2008 and 2017 and analyzed their diagnostic pathway, cancer stage at diagnosis, and survival time. Results: Of the study participants, 59,423 patients (about 15.3%) were diagnosed with cancer through the emergency department. We found that a sizable proportion of older people and patients with low education and low incomes were diagnosed through emergency department visits, and those with a health condition comorbidity severity of 3 had the highest proportion diagnosed by the emergency department, advanced stages at diagnosis, and risk of death. These can be classified as high-risk groups. In addition, 76.4% of patients diagnosed in the emergency department had advanced cancer, and the risk of death was 1.46 times higher than that of patients diagnosed in the non-emergency department. Although cancer screening is available, it does not reduce the proportion of patients with advanced cancer who are diagnosed through or at the time of diagnosis in the emergency department. Conclusions: The present study found that the government's cancer screening did not affect the proportion or number of cancers diagnosed through emergency department visits. Therefore, the government should focus on more cancer screening, health education in high-risk groups, and strengthening the link between emergency and oncology departments to reduce the risk of death for patients diagnosed through emergency department visits.

Impact of Time Period and Birth Cohort on the Trend of Advanced Neoplasm Prevalence in the 40-49 Average-Risk Screening Population.

BACKGROUND AND AIMS: Early-onset colorectal cancer (CRC) is increasing globally. While the United States have lowered the age of initiation of screening to 45 years, other countries still start screening at 50 years of age. In Taiwan, the incidence of CRC has declined in 55- to 74-year-olds after the initiation of screening, but still increased in those 50-54 years of age, potentially due to rising precancerous lesion incidence in 40- to 49-year-olds. This study aimed to explore the chronological trend of the prevalence of colorectal advanced neoplasms (AN) in the screening population 40-54 years of age. METHODS: We retrospectively analyzed a screening colonoscopy cohort for prevalence of AN in average-risk subjects 40-54 years of age from 2003 to 2019. Logistic regression was used to distinguish cohort effect from time-period effect on the prevalence of AN. RESULTS: In total, 27,805 subjects (52.1% male) men were enrolled. There were notable increases in prevalence of AN in all 3 age groups during the 17-year span, but these were more rapid in those 40-44 years of age (0.99% to 3.22%) and 45-49 years of age (2.50% to 4.19%). Those 50-54 years of age had a higher risk of AN (adjusted odds ratio [aOR], 1.62; 95% confidence interval [CI], 1.19-2.19) in 2003-2008 but not in later periods (2009-2014: aOR, 1.08; 95% CI, 0.83-1.41; 2015-2019: aOR, 0.76; 95% CI, 0.56-1.03) when compared with those 45-49 years of age. CONCLUSION: The prevalence of AN in those 40-54 years of age increased in the Taiwanese population, with a later birth cohort having a higher prevalence of AN. However, the prevalence of AN in those 45-49 years of age increased more remarkably and approximated that in those 50-54 years of age, which may justify earlier initiation of CRC screening in those 45 years of age.

Response of bacterial and fungal communities in natural biofilms to bioavailable heavy metals in a mining-affected river.

Biofilms, known as "microbial skin" in rivers, respond to rapid and sensitive environmental changes. However, the ecological response mechanisms of bacterial and fungal communities in river biofilms toward heavy metal pollution (HMP) remains poorly understood. This study focused on the key driving factors of bacterial and fungal community diversity and composition and their ecological response mechanisms within periphytic biofilms of Asia's largest Pb-Zn mining area. The diversity, dominant bacterial taxa, and bacteria structure in biofilms were influenced by biologically available heavy metal (HM) fractions, with Ni-F3 (17.96 %) and Pb-F4 (16.27 %) as the main factors affecting the bacterial community structure. Fungal community structure and α-diversity were more susceptible to physicochemical parameters (pH and nutrient elements). Partial least squares path modeling revealed that environmental factors influencing bacterial and fungal communities in biofilms were ranked as water quality > metal fractions > total metals. Dispersal limitation was the most critical ecological process in bacterial (56.9 %) and fungal (73.4 %) assembly. The proportion of heterogeneous selection by bacteria (39.5 %) was higher than that of fungus (26.2 %), which increased with increasing HMP. Bacterial communities had a higher migration rate (0.48) and ecological drift proportion (3.6 %), making them more prone to escape environmental stress. Fungal communities exhibited more keystone species, larger niche width (23.24 ± 13.04 vs. 9.72 ± 5.48), higher organization level, and a more stable co-occurrence network than bacterial communities, which enabled them to better adapt to high environmental pollution levels. These findings expanded the understanding of the spatiotemporal dynamics of microbial communities within biofilms in HM-polluted watersheds and provided new insights into the ecological responses of microbial communities to HMP.

Biofilm-mediated heavy metal bioaccumulation and trophic transfer in a mining-contaminated river.

Biofilms, essential for material circulation and energy flow in aquatic ecosystems, markedly enrich heavy metals in water environments. However, the impact of these accumulated metals on organisms feeding on biofilms remains poorly unknown. This study involved a year-long seasonal survey along the Bijiang River, located next to Asia's largest lead (Pb)-zinc (Zn) mine, conducted to investigate the role of biofilms in nutrient and metal transfer in food webs. In total, 355 biotic and abiotic samples, including water, biofilms, and aquatic biota, were analyzed for the presence of eight heavy metals (arsenic [As], cadmium, chromium, copper, Pb, nickel, Zn, and iron) as well as stable carbon (δ13C) and nitrogen (δ15N) isotopes. Wide ranges of δ13C and δ15N values indicated diverse dietary carbon sources and trophic positions in the Bijiang River (maximum trophic level: 4.28). A Bayesian mixing model revealed that periphytic biofilms were the dominant basal carbon source, especially in spring, whereas in summer, consumers exploited more diverse food sources, possibly because feeding on spring biofilms enhanced predator feeding efficiency. Metals tended to be biodiluted along food chains owing to their higher concentrations in biofilms and benthic organisms as well as their chemical forms. Although diet did not significantly affect heavy metal accumulation in fish, those relying on biofilms as the main carbon source showed significantly higher As (p = 0.048) and Pb (p = 0.007) levels compared with those relying on C4 plants. Overall, this study highlights the critical role of periphytic biofilms in nutrient and metal dynamics in aquatic food webs.

Implementation of Accurate Parameter Identification for Proton Exchange Membrane Fuel Cells and Photovoltaic Cells Based on Improved Honey Badger Algorithm.

Predicting the system efficiency of green energy and developing forward-looking power technologies are key points to accelerating the global energy transition. This research focuses on optimizing the parameters of proton exchange membrane fuel cells (PEMFCs) and photovoltaic (PV) cells using the honey badger algorithm (HBA), a swarm intelligence algorithm, to accurately present the performance characteristics and efficiency of the systems. Although the HBA has a fast search speed, it was found that the algorithm's search stability is relatively low. Therefore, this study also enhances the HBA's global search capability through the rapid iterative characteristics of spiral search. This method will effectively expand the algorithm's functional search range in a multidimensional and complex solution space. Additionally, the introduction of a sigmoid function will smoothen the algorithm's exploration and exploitation mechanisms. To test the robustness of the proposed methodology, an extensive test was conducted using the CEC'17 benchmark functions set and real-life applications of PEMFC and PV cells. The results of the aforementioned test proved that with regard to the optimization of PEMFC and PV cell parameters, the improved HBA is significantly advantageous to the original in terms of both solving capability and speed. The results of this research study not only make definite progress in the field of bio-inspired computing but, more importantly, provide a rapid and accurate method for predicting the maximum power point for fuel cells and photovoltaic cells, offering a more efficient and intelligent solution for green energy.

Investigation of Biomechanical Differences in Level Walking between Patients with Bilateral and Unilateral Total Knee Replacements.

Due to the high risk of a bilateral total knee arthroplasty (TKR) following unilateral TKR, this study was performed to investigate bilateral TKR patients. Specifically, we examined biomechanical differences between the first replaced and second replaced limbs of bilateral patients. Furthermore, we examined bilateral TKR effects on hip, knee, and ankle biomechanics, compared to the replaced and non-replaced limbs of unilateral patients. Eleven bilateral patients (70.09 ± 5.41 years, 1.71 ± 0.08 m, 91.78 ± 13.00 kg) and fifteen unilateral TKR patients (65.67 ± 6.18 years, 1.73 ± 0.10 m, 87.72 ± 15.70 kg) were analyzed while performing level walking. A repeated measures one-way ANOVA was performed to analyze between-limb differences within the bilateral TKR group. A 2 × 2 (limb × group) ANOVA was used to determine differences between bilateral and unilateral patients. Our results showed that the second replaced limb exhibited a lower peak initial-stance knee extension moment than the first replaced limb. No other kinematic or kinetic differences were found. Bilateral patients exhibited lower initial-stance knee extension moments, knee abduction moments, and dorsiflexion moments, compared to unilateral patients. Bilateral patients also exhibited lower push-off peak hip flexion moments and vertical GRF. The differences between the first and second replaced limbs of bilateral patients may indicate different adaptation strategies used following a second TKR. The significant group differences indicate that adaptations are different between these groups, and it is not recommended to use patients with unilateral and bilateral TKR together in gait analyses.

Combined Effect of Hydrophilic Pore and the Type of Protons on Proton Conductivity in Porous Metal-Organic Frameworks: A Feasible Approach to Achieve a Super Proton Conductor under Hydrated Conditions.

There has been a steady growth of interest in proton-conductive metal-organic frameworks (MOFs) due to their potential utility in proton-exchange membrane fuel cells. To attain a super proton conductivity (>1 × 10-2 S cm-1) in a MOF-based proton conductor is a key step toward practical application. Currently, most studies are focused on enhancing the proton conductivity of porous MOFs by controlling a single factor, such as the type of protons or hydrophilic pore or hydrogen bond. However, a limited contribution from a single factor cannot afford to remarkably increase the proton conductivity of the MOF and form a super proton conductor. Herein, we constructed two distinct porous MOFs, {(H3O+)4[Cu12(ci)12(OH)4(H2O)12]·3H2O·9DMF} (Cu-ci-3D, H2ci = 1H-indazole-5-carboxylic acid, DMF = N,N'-dimethylformamide) and {[Co(Hppca)2]·2HN(CH3)2·CH3OH·2H2O} (Co-ppca-2D, H2ppca = 5-(pyridin-3-yl)-1H-pyrazole-3-carboxylic acid), to tune their proton conductivities at high relative humidity (RH) using the combined effect of hydrophilic pore and the type of protons, ultimately achieving super proton conduction. Excitingly, Cu-ci-3D indeed harvests a super proton conductivity of 1.37 × 10-2 S cm-1 at 353 K and ∼97% RH, superior to some previously reported MOF-based proton conductors. The results present a unique perspective for developing high-performance MOF-based proton conductors and understanding their structure-performance relationships.

Quantification of 3-Dimensional Confluence-Atrial Morphology in Supracardiac Total Anomalous Pulmonary Venous Connection.

Background: Pulmonary vein stenosis (PVS) continues to be a major complication after surgical repair of total anomalous pulmonary venous connection (TAPVC). Recent studies suggest that the morphology of pulmonary venous confluence and the left atrium (LA) is associated with PVS. However, there are limited data on the prognostic value of integrating quantitative confluence-atrial morphology into risk stratification. Objectives: This study sought to evaluate the prognostic impact of novel imaging metrics derived from 3-dimensional (3D) computed tomography angiography (CTA) modeling on postsurgical PVS (PPVS) in the supracardiac TAPVC (sTAPVC) setting. Methods: Patients undergoing sTAPVC repair in 2017 to 2022 from 3 centers were retrospectively reviewed. Study investigators developed 3D CTA modeled geometric features to quantify confluence-atrial morphology that were analyzed with regard to PPVS. Results: Of the 162 patients (median age 61 days; 55% having preoperative pulmonary venous obstruction [prePVO]) included, 47 (29%) with PPVS at a median of 1.5 months ([quartile 1-quartile 3: 1.5-3.0 months]). In the univariable analysis, the indexed total volume of the LA and confluence (iTVLC) and the ratio of the corresponding confluence length to the mean distance between the LA and confluence (CCL/mDBLC ratio) were significantly associated with PPVS. In a multivariable model adjusting for prePVO and age, the iTVLC and CCL/mDBLC ratio independently predicted PPVS (HR: 1.15; 95% CI: 1.06-1.25; and HR: 1.20; 95% CI: 1.08-1.35, respectively, all P < 0.01). Specifically, an iTVLC ≥20 cm3/m2 and a CCL/mDBLC ratio ≥7.7 were significantly associated with a reduced risk of PPVS. Conclusions: Quantification of 3D confluence-atrial morphology appears to offer a deeper and better metric to predict PPVS in patients with sTAPVC.

Assessing CO2 sources and sinks in and around Taiwan: Implication for achieving regional carbon neutrality by 2050.

Taiwan has pledged to achieve net-zero carbon emissions by 2050, but the current extent of carbon sinks in Taiwan remains unclear. Therefore, this study aims to first review the existing nature-based carbon sinks on land and in the oceans around Taiwan. Subsequently, we suggest potential strategies to reduce CO2 emissions and propose carbon dioxide removal methods (CDRs). The natural carbon sinks by forests, sediments, and oceans in and around Taiwan are approximately 21.5, 42.1, and 96.8 Mt-CO2 y-1, respectively, which is significantly less than Taiwan's CO2 emissions (280 Mt-CO2 y-1). Taiwan must consider decarbonization strategies like using electric vehicles, renewable energy, and hydrogen energy by formulating enabling policies. Besides more precisely assessing both terrestrial and marine carbon sinks, Taiwan should develop novel CDRs such as bioenergy with carbon capture and storage, afforestation, reforestation, biochar, seaweed cultivation, and ocean alkalinity enhancement, to reach carbon neutrality by 2050.

The impact of flooding from the Minjiang River on the succession of harmful algal blooms (HABs) caused by diatoms in China's offshore waters.

This study examines diatom assemblages in the Matsu Archipelago, an area influenced by Minjiang River runoff. It focuses on harmful algal blooms (HABs) that occurred between August 2021 and July 2022. Utilizing 18S rRNA metabarcoding and microscopic analysis, we observed a significant diatom bloom during early summer runoff, peaking at 5 × 105 cells L-1. The research reveals dynamic community changes during the runoff season, with dominant genera including Pseudo-nitzschia, Chaetoceros, and Skeletonema. Skeletonema cell density correlated with NO3 levels, Chaetoceros had a slight PO4 affinity, and Pseudo-nitzschia showed a negative correlation with Skeletonema. Pseudo-nitzschia, which prefers high light and pH conditions, had notably high concentrations in the flood season and in the autumn. In both, it was dominated by potential toxin-producing species - P. multistriata and P. pungens during the flooding, and P. cuspidate in the autumn. These findings highlight the intricate relationship between diatom dynamics and environmental factors, providing essential insights for managing HABs, especially Pseudo-nitzschia species, amidst environmental changes.

Contribution of greenness, air pollution, and residential food environment to excess gestational weight gain: A cross-sectional study in Wuhan, China.

OBJECTIVES: Our goal was to explore how greenness, air pollution, and residential food environment were linked to excessive gestational weight gain (EGWG), and to estimate their combined effects on this condition. METHOD: This cross-sectional analysis included 51,507 pregnant women from the Wuhan Maternal and Child Health Management Information System between 2016 and 2019. Generalized linear mixed regression models were employed to explore the relationships between greenness, air pollution, residential food environmental exposure, and EGWG; and the combined effects were further estimated by cluster analysis and principal components analysis. RESULT: We only found a significant association between convenience store density within the 250 m buffer zone (OR = 1.03 and 95% CI: 1.01,1.05) and EGWG. In terms of air pollution, sulfur dioxide(SO2), particulate matter with a diameter of 10 µm or less(PM10), and particulate matter with a diameter of 2.5 µm or less(PM2.5) were substantially correlated with a higher prevalence of EGWG and higher GWG, with (OR = 1.16 and 95% CI: 1.12,1.21; OR = 1.12 and 95% CI: 1.08,1.16; OR = 1.17 and 95% CI: 1.14,1.21, respectively) per interquartile range(IQR) increase. Cluster analysis revealed the presence of three clusters representing urban exposures. In contrast to urban environment clusters characterized by favourable conditions, those exhibiting elevated air pollution levels, high-density residential food environment and low levels of greenness were found to have increased odds of EGWG (OR = 1.10, 95% CI: 1.03, 1.19). CONCLUSION: This study emphasizes that exposure to elevated air pollution, high-density residential neighbourhood food environments, and low levels of greenness is a neighbourhood obesogenic environment for pregnant women.

Nutrient sources, phytoplankton blooms, and hypoxia along the Chinese coast in the East China Sea: Insight from 2014.

Phytoplankton blooms are common along the Chinese coast in the East China Sea, driven by various nutrient sources including river discharge, bottom water regeneration, and Kuroshio subsurface water intrusion. A notable 2014 summer bloom off the Zhejiang coast, exhibiting a Chl a concentration of 20.1 µg L-1, was significantly influenced by Changjiang River discharge, and high nutrient concentrations are often observed in the region's surface water. During blooms, primary production peaks at 1686.3 mg C m-3 d-1, indicating substantial CO2 absorption, with surface water fCO2 declining to 299.5 µatm, closely linked to plankton activities. Hypoxia often coincides with these frequent bloom occurrences, implicating marine-derived organic matter decomposition as a pivotal factor. Elevated particulate organic carbon concentrations further support this assumption, alongside increased nutrient levels, fCO2, and low pH in hypoxic waters. These findings underscore the intricate interplay between phytoplankton, nutrient cycling, and hypoxia formation, essential for effective coastal ecosystem management.

Fabrication electro-spun Poly(vinyl alcohol)-Melamine nonwoven membrane composite separator for high-power lithium-ion batteries.

Current commercial separators used in lithium-ion batteries have inherent flaws, especially poor thermal stability, which pose substantial safety risks. This study introduces a high-safety composite membrane made from electrospun poly(vinyl alcohol)-melamine (PVAM) and polyvinylidene fluoride (PVDF) polymer solutions via a dip coating method, designed for high-voltage battery systems. The poly(vinyl alcohol) and melamine components enhance battery safety, while the PVDF coating improves lithium-ion conductivity. The dip-coated PVDF/Esp-PVAM composite separators were evaluated for electrolyte uptake, contact angle, thermal stability, porosity, electrochemical stability and ionic conductivity. Notably, our Dip 1 % PVDF@Esp-PVAM composite separator exhibited excellent wettability and a lithium-ion conductivity of approximately 7.75 × 10⁻4 S cm⁻1 at room temperature. These separators outperformed conventional PE separators in half-cells with Ni-rich NCM811 cathodes, showing exceptional cycling stability with 93.4 % capacity retention after 100 cycles at 1C/1C, as compared to 84.8 % for PE separators. Our Dip 1 % PVDF@Esp-PVAM composite separator demonstrates significant potential for enhancing the long-term durability and high-rate performance of lithium-ion batteries, making it a promising option for long-term energy storage applications.

The survival and cost-effectiveness analysis of adjunctive Chinese medicine therapy for patients with non-small cell lung cancer: a nationwide cohort study in Taiwan.

Introduction: Cancer, particularly lung cancer, is a significant global healthcare challenge. Non-Small Cell Lung Cancer (NSCLC) constitutes 85% of cases. Patients often seek alternative therapies like Chinese medicine alongside Western treatments. This study investigates the survival outcomes and cost-effectiveness of adjunctive Chinese medicine therapy for NSCLC patients in Taiwan. Methods: We utilized the National Health Insurance Research Database in a retrospective cohort study from 2000 to 2018, focusing on NSCLC patients diagnosed between 2007 and 2013. After propensity score matching 1:5 ratio, then compared patients with and without adjunctive Chinese medicine therapy. Survival outcomes, cost-effectiveness, and sensitivity analyses were conducted. Results: The study involved 43,122 NSCLC patients with 5.76% receiving adjunctive Chinese medicine. There is no significant associated between the risk of death and adjuvant Chinese medicine therapy until 181-365 days of adjuvant treatment could reduce the risk of death (HR = 0.88, 95% CI: 0.80-0.98). Cost-effectiveness analysis showed an incremental cost-effectiveness ratio of 880,908 NT$/year. Conclusion: Adjunctive Chinese medicine therapy, particularly when administered for 181-365 days, significantly reduced the mortality risk among stage IV NSCLC patients. The cost-effectiveness aligns with willingness-to-pay thresholds, indicating economic benefit.

Targeted protein degradation systems to enhance Wnt signaling.

Molecules that facilitate targeted protein degradation (TPD) offer great promise as novel therapeutics. The human hepatic lectin asialoglycoprotein receptor (ASGR) is selectively expressed on hepatocytes. We have previously engineered an anti-ASGR1 antibody-mutant RSPO2 (RSPO2RA) fusion protein (called SWEETS) to drive tissue-specific degradation of ZNRF3/RNF43 E3 ubiquitin ligases, which achieved hepatocyte-specific enhanced Wnt signaling, proliferation, and restored liver function in mouse models, and an antibody-RSPO2RA fusion molecule is currently in human clinical trials. In the current study, we identified two new ASGR1- and ASGR1/2-specific antibodies, 8M24 and 8G8. High-resolution crystal structures of ASGR1:8M24 and ASGR2:8G8 complexes revealed that these antibodies bind to distinct epitopes on opposing sides of ASGR, away from the substrate-binding site. Both antibodies enhanced Wnt activity when assembled as SWEETS molecules with RSPO2RA through specific effects sequestering E3 ligases. In addition, 8M24-RSPO2RA and 8G8-RSPO2RA efficiently downregulate ASGR1 through TPD mechanisms. These results demonstrate the possibility of combining different therapeutic effects and degradation mechanisms in a single molecule.

Design principles and therapeutic applications of novel synthetic WNT signaling agonists.

Wingless-related integration site or Wingless and Int-1 or Wingless-Int (WNT) signaling is crucial for embryonic development, and adult tissue homeostasis and regeneration, through its essential roles in cell fate, patterning, and stem cell regulation. The biophysical characteristics of WNT ligands have hindered efforts to interrogate ligand activity in vivo and prevented their development as therapeutics. Recent breakthroughs have enabled the generation of synthetic WNT signaling molecules that possess characteristics of natural ligands and potently activate the pathway, while also providing distinct advantages for therapeutic development and manufacturing. This review provides a detailed discussion of the protein engineering of these molecular platforms for WNT signaling agonism. We discuss the importance of WNT signaling in several organs and share insights from the initial application of these new classes of molecules in vitro and in vivo. These molecules offer a unique opportunity to enhance our understanding of how WNT signaling agonism promotes tissue repair, enabling targeted development of tailored therapeutics.