Xu Ma

Periconceptional folic acid supplementation (FAS) is known to reduce the risk of adverse pregnancy outcomes, but limited evidence exists on its national co

Maple syrup urine disease (MSUD) is an inherited metabolic disorder caused by a deficiency in the activity of the hepatic branched-chain α-ketoacid dehydrogenase (BCKDH) complex, which leads to the toxic accumulation of three branched-chain amino acids (BCAAs) and their respective α-ketoacid, resulting in severe neurotoxicity, coma and even death without effective therapeutic measures. In this study, we established the patient induced pluripotent stem cells (iPSC)-derived hepatic organoids (HOs), analyzed the characteristics, and applied adenine base editor (ABE8e) to correct a mutation (T322I) of the BCKDHB (branched chain keto acid dehydrogenase E1, beta polypeptide) gene in patient induced pluripotent stem cells (iPSC)-derived hepatic organoids (HOs). qRT-PCR and western blot analysis were performed to assess the expression level of BCKDHA (branched chain keto acid dehydrogenase E1, alpha polypeptide) and BCKDHB. The effects of base editing were comprehensively analyzed using both bulk RNA sequencing and single-cell RNA sequencing (scRNA-Seq). Immunofluorescence and RT-PCR arrayed the high expression of hepatoblast specific proteins in HOs, such as α-1-anti-trypsin (A1AT), hepatocyte nuclear factor-4-alpha (HNF4A), cytokeratin18 (CK18), albumin (ALB), cytochrome P450 family 3 subfamily A member 4 (CYP3A4) and cytochrome P450 family 3 subfamily A member 7(CYP3A7). Functional experiments indicated that these HOs recapitulated characteristics of hepatocytes like glycogen accumulation, low-density lipoprotein (LDL) uptake, indocyanine green (ICG) uptake and release as well as quantitation of ALB and urea from HOs. The levels of BCKDHA and BCKDHB were dramatically decreased in MSUD-HOs compared with control-HOs (P < 0.01) detected by qRT-PCR, western blot and immunofluorescence. Deep sequencing and whole genome sequencing (WGS) demonstrated that the correction of BCKDHB mutation in patient iPSC-derived HOs caused high on-target gene editing without any detectable off-target effects. Moreover, the corrected MSUD-HOs exhibited restored BCKDH enzymatic function and reduced BCAAs level. The transcriptome analysis indicates that the MSUD-HOs with BCKDHB mutation reduced mRNA level of regulating metabolism associated with liver mitochondrial function, while the corrected MSUD-HOs rescued those processes after ABE8e correction. The scRNA-Seq analysis further validated the rescue effects of BCKDH function after gene editing. Our study provides reliable evidence that ABE8e is highly efficient and safe in correcting patient-derived HOs from MSUD, indicating the feasibility to be a transformative treatment for genetic hepatic diseases like MSUD.

The development of gene editing tools such as CRISPR-Cas9 and base editors (BE) is critical for genetic diseases and cancer. Lentivirus-like particles (LVLPs) grows into an auspicious platform for delivering mRNA or ribonucleic proteins (RNPs) due to it integrates the advantage of viral and non-viral vectors. Current LVLP systems predominantly utilize HIV-Gag and Pol proteins. However, the reverse transcriptase and integrase of Pol, pose risks of genomic integration and potential tumorigenesis. Enhancing the safety of VLP system is essential. This study focuses on improving the LVLP to minimize these risks. We implemented a Gag-Only strategy, constructing LVLPs with HIV-Gag protein, thereby eliminating the integration risks linked to Pol. By leveraging the interactions between MS2-MCP (MS2 coat protein), PP7 and PP7 BP (PP7 binding protein), and the psi (HIV packaging signal) with HIV-Gag, we encapsulated PAMless andesine base editor (CE-8e-SpRY) mRNA and sgRNA targeting the PD1 start codon (ATG) into the LVLP. Using recombinant lentiviral vector technology, we developed a stable PD1-expressing 293T cell line (PD1-293T) to assess the editing efficiency of LVLP. The psi-LVLP demonstrated effective packaging capabilities, achieving 15% base editing efficiency in 293T cells. By optimizing plasmid ratios, we observed increased CE-8e-SpRY mRNA copy numbers, with 30% base editing efficiency. Additionally, the integration of HDVrz (hepatitis delta virus ribozyme) and psi into sgRNA (HDVrz-psi-LVLP) substantially enhanced sgRNA copy numbers, resulting in approximately 50% base editing efficiency in 293T cells and 20% base editing efficiency in Jurkat cells. Mendelian randomization analyses revealed significant genetic correlations between PD1, B2M, CIITA, and TIGIT genes with various cancer risks. Furthermore, HDVrz-psi-LVLPs targeting the start codons of B2M, CIITA, and TIGIT exhibited high base editing activity in both Jurkat and 293T cells. In conclusion, this optimized platform effectively encapsulates CE-8e-SpRY mRNA and sgRNA, achieving high editing efficiencies across multiple genes and cell types. We introduce a safer and more efficient gene editing tool delivery system by constructing LVLPs based on the Gag-Only strategy. Our study presents a promising implication for cancer immunotherapy.

Climate change may affect fertility; however, associations between ambient temperature and human fecundity remain understudied. We assessed the relationship between temperature and fecundity in 2,443,879 reproductive-age couples from 350 cities across China, with follow-up every 3 months until conception or for up to 1 year. We used Cox regression models to evaluate associations of 1-year, 6-month, and 3-month average temperature exposures preceding conception or censoring with fecundability odds ratios (FORs). Accelerated failure time and multivariate linear regression models were applied to examine associations between temperature and time to pregnancy (TTP). During the one-year follow-up, 2,038,378 (83.4%) couples conceived. Higher temperatures were associated with reduced fecundity and extended TTP, with monotonic exposure-response curves observed. Specifically, a 1°C increase in 1-year average temperature was associated with a 4.2% decrease in fecundity (FOR=0.958, 95% confidence interval [CI]: 0.957, 0.959). The corresponding absolute probability of pregnancy within 1 year decreased by 1.35% (absolute risk difference=–1.35%, 95% CI: –1.41%, –1.26%). Each 1°C increase in annual average temperature was associated with a 2.5% increase in median TTP for all couples (time ratio=1.025, 95% CI: 1.024, 1.025). Among couples who conceived within 1 year, TTP was prolonged by 1.60% (95% CI: 1.55%, 1.65%). Smaller FOR estimates, indicating reduced fecundity, were observed in the East, South, and North regions; in tropical and temperate monsoon zones; and among couples where either partner was overweight/obese, woman was attempting her first pregnancy or exposed to second-hand smoke, or man consumed alcohol. Increased temperatures may adversely impact fecundity by prolonging TTP, highlighting the need for heat mitigation strategies.

Objective: Evidence on the associations between fine particulate matter (PM2.5) and diabetes risk from women of reproductive age, on whom diabetes may have adverse long-term health effects on both themselves and future generations, remains scarce. We therefore examined the associations of long-term PM2.5 exposure with fasting blood glucose (FBG) levels and diabetes risk among reproductive-aged women in China.Research Design and Methods: This study included 20,076,032 women aged 20-49 years participating in the National Free Preconception Health Examination Project in China between 2010 and 2015. PM2.5 was estimated utilizing a satellite-based model. Multivariate linear and logistic regression models were used to examine the associations of PM2.5 exposure with FBG level and diabetes risk, respectively. Diabetes burden attributable to PM2.5 was estimated using attributable fraction (AF) and attributable number.Results: PM2.5 showed monotonic relationships with elevated FBG level and diabetes risk. Each interquartile range (IQR, 27 μg/m3) increase in 3-year average PM2.5 concentration was associated with 0.078 mmol/L (95% confidence interval [CI]: 0.077, 0.079) increase of FBG and 18% (95%CI: 16%, 19%) higher risk of diabetes. The AF attributed to PM2.5 exposure exceeding 5 μg/m3 was 29.0% (95%CI: 27.5%, 30.5%), corresponding to an additional 78.6 (95%CI: 74.5, 82.6) thousand diabetes cases. Subgroup analyses showed more pronounced diabetes risks among those who were overweight or obese, above 35 years, less educated, of minority ethnicity, with rural household registration, and residing in Western China.Conclusions: We found long-term PM2.5 exposure was associated with higher diabetes risk among women of reproductive age in China.

Objective: Evidence on the associations between fine particulate matter (PM2.5) and diabetes risk from women of reproductive age, on whom diabetes may have adverse long-term health effects on both themselves and future generations, remains scarce. We therefore examined the associations of long-term PM2.5 exposure with fasting blood glucose (FBG) levels and diabetes risk among reproductive-aged women in China.Research Design and Methods: This study included 20,076,032 women aged 20-49 years participating in the National Free Preconception Health Examination Project in China between 2010 and 2015. PM2.5 was estimated utilizing a satellite-based model. Multivariate linear and logistic regression models were used to examine the associations of PM2.5 exposure with FBG level and diabetes risk, respectively. Diabetes burden attributable to PM2.5 was estimated using attributable fraction (AF) and attributable number.Results: PM2.5 showed monotonic relationships with elevated FBG level and diabetes risk. Each interquartile range (IQR, 27 μg/m3) increase in 3-year average PM2.5 concentration was associated with 0.078 mmol/L (95% confidence interval [CI]: 0.077, 0.079) increase of FBG and 18% (95%CI: 16%, 19%) higher risk of diabetes. The AF attributed to PM2.5 exposure exceeding 5 μg/m3 was 29.0% (95%CI: 27.5%, 30.5%), corresponding to an additional 78.6 (95%CI: 74.5, 82.6) thousand diabetes cases. Subgroup analyses showed more pronounced diabetes risks among those who were overweight or obese, above 35 years, less educated, of minority ethnicity, with rural household registration, and residing in Western China.Conclusions: We found long-term PM2.5 exposure was associated with higher diabetes risk among women of reproductive age in China.

OBJECTIVE: Evidence of the associations between fine particulate matter (PM2.5) and diabetes risk from women of reproductive age, in whom diabetes may have adverse long-term health effects for both themselves and future generations, remains scarce. We therefore examined the associations of long-term PM2.5 exposure with fasting blood glucose (FBG) level and diabetes risk in women of reproductive age in China. RESEARCH DESIGN AND METHODS: This study included 20,076,032 women age 20–49 years participating in the National Free Preconception Health Examination Project in China between 2010 and 2015. PM2.5 was estimated using a satellite-based model. Multivariate linear and logistic regression models were used to examine the associations of PM2.5 exposure with FBG level and diabetes risk, respectively. Diabetes burden attributable to PM2.5 was estimated using attributable fraction (AF) and attributable number. RESULTS: PM2.5 showed monotonic relationships with elevated FBG level and diabetes risk. Each interquartile range (27 μg/m3) increase in 3-year average PM2.5 concentration was associated with a 0.078 mmol/L (95% CI 0.077, 0.079) increase in FBG and 18% (95% CI 16%, 19%) higher risk of diabetes. The AF attributed to PM2.5 exposure exceeding 5 μg/m3 was 29.0% (95% CI 27.5%, 30.5%), corresponding to an additional 78.6 thousand (95% CI 74.5, 82.6) diabetes cases. Subgroup analyses showed more pronounced diabetes risks in those who were overweight or obese, age >35 years, less educated, of minority ethnicity, registered as a rural household, and residing in western China. CONCLUSIONS: We found long-term PM2.5 exposure was associated with higher diabetes risk in women of reproductive age in China.