Type 1 diabetes mellitus (T1DM) is the most common chronic autoimmune disease in young patients and is characterized by the loss of pancreatic beta cells; as a result, the body becomes insulin ...deficient and hyperglycemic. Administration or injection of exogenous insulin cannot mimic the endogenous insulin secreted by a healthy pancreas. Pancreas and islet transplantation have emerged as promising treatments for reconstructing the normal regulation of blood glucose in T1DM patients. However, a critical shortage of pancreases and islets derived from human organ donors, complications associated with transplantations, high cost, and limited procedural availability remain bottlenecks in the widespread application of these strategies. Attempts have been directed to accommodate the increasing population of patients with T1DM. Stem cell therapy holds great potential for curing patients with T1DM. With the advent of research on stem cell therapy for various diseases, breakthroughs in stem cell-based therapy for T1DM have been reported. However, many unsolved issues need to be addressed before stem cell therapy will be clinically feasible for diabetic patients. In this review, we discuss the current research advances in strategies to obtain insulin-producing cells (IPCs) from different precursor cells and in stem cell-based therapies for diabetes. Keywords: Type 1 diabetes mellitus, Stem cells, Insulin-producing cells, Pancreatic islets, Transplantation
A comprehensive cellular anatomy of normal human kidney is crucial to address the cellular origins of renal disease and renal cancer. Some kidney diseases may be cell type-specific, especially renal ...tubular cells. To investigate the classification and transcriptomic information of the human kidney, we rapidly obtained a single-cell suspension of the kidney and conducted single-cell RNA sequencing (scRNA-seq). Here, we present the scRNA-seq data of 23,366 high-quality cells from the kidneys of three human donors. In this dataset, we show 10 clusters of normal human renal cells. Due to the high quality of single-cell transcriptomic information, proximal tubule (PT) cells were classified into three subtypes and collecting ducts cells into two subtypes. Collectively, our data provide a reliable reference for studies on renal cell biology and kidney disease.
To identify accessible and permissive human cell types for efficient derivation of induced pluripotent stem cells （iPSCs）, we investigated epigenetic and gene expression signatures of multiple ...postnatal cell types such as fibroblasts and blood cells. Our analysis suggested that newborn cord blood （CB） and adult peripheral blood （PB） mononuclear cells （MNCs） display unique signatures that are closer to iPSCs and human embryonic stem cells （ESCs） than agematched fibroblasts to iPSCs/ESCs, thus making blood MNCs an attractive cell choice for the generation of integration-free iPSCs. Using an improved EBNA1/OriP plasmid expressing 5 reprogramming factors, we demonstrated highly efficient reprogramming of briefly cultured blood MNCs. Within 14 days of one-time transfection by one plasmid, up to 1000 iPSC-like colonies per 2 million transfected CB MNCs were generated. The efficiency of deriving iPSCs from adult PB MNCs was approximately 50-fold lower, but could be enhanced by inclusion of a second EBNA1/ OriP plasmid for transient expression of additional genes such as SV40 T antigen. The duration of obtaining bona fide iPSC colonies from adult PB MNCs was reduced to half （-14 days） as compared to adult fibroblastic cells （28- 30 days）. More than 9 human iPSC lines derived from PB or CB blood cells are extensively characterized, including those from PB MNCs of an adult patient with sickle cell disease. They lack V（D）J DNA rearrangements and vector DNA after expansion for 10-12 passages. This facile method of generating integration-free human iPSCs from blood MNCs will accelerate their use in both research and future clinical applications.
Dichlorodiphenyltrichloroethane (DDT) is well known for its harmful effects and has been banned around the world. However, DDT is still frequently detected in natural environments, particularly in ...aquaculture and harbor sediments. In this study, 15 surface sediment samples were collected from a typical tropical bay (Zhanjiang Bay) in the South China Sea, and the levels of DDT and its metabolites in sediment and porewater samples were investigated. The results showed that concentrations of DDXs (i.e., DDT and its metabolites) in bulk sediments were 1.58–51.0 ng g−1 (mean, 11.5 ng g−1). DDTs (DDT and its primary metabolites, dichlorodiphenyldichloroethane (DDD) and dichlorodiphenyldichloroethylene (DDE)) were the most prominent, accounting for 73.2%–98.3% (86.1% ± 12.8%) of the DDXs. Additionally, high-order metabolites (i.e., 1-chloro-2,2-bis(4′-chlorophenyl)ethylene (p,p′-DDMU), 2,2-bis(p-chlorophenyl)ethylene (p,p′-DDNU), 2,2-bis(p-chlorophenyl)ethanol (p,p′-DDOH), 2,2-bis(p-chlorophenyl)methane (p,p′-DDM), and 4,4′-dichlorobenzophenone (p,p′-DBP)) were also detected in most of the sediment and porewater samples, with DDMU and DBP being predominant. The DDTs concentration differed among the sampling sites, with relatively high DDTs concentrations in the samples from the aquaculture zone and an area near the shipping channel and the Haibin shipyard. The DDD/DDE ratios indicated a reductive dichlorination of DDT to DDD under anaerobic conditions at most of the sampling sites of Zhanjiang Bay. The possible DDT degradation pathway in the surface sediments of Zhanjiang Bay was p,p′-DDT/p,p′-DDD(p,p′-DDE)/p,p′-DDMU/p,p′-DDNU/ … /p,p′-DBP. The DDXs in the sediments of Zhanjiang Bay were mainly introduced via mixed sources of industrial DDT and dicofol, including fresh input and historical residue. The concentrations of DDXs in porewater samples varied from 66.3 to 250 ng L−1, exhibiting a distribution similar to that in the accompanying sediments. However, the content of high-order metabolites was relatively lower in porewater than in sediment, indicating that high-order degradation mainly occurs in particles. Overall, this study helps in understanding the distribution, source, and degradation of DDT in a typical tropical bay.
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•DDTs are still frequently detected in sediment/porewater of a typical tropical bay.•High-order metabolites of DDXs were detected in most sediment and porewater samples.•DDTs were derived from mixed sources including fresh input and historical residue.•DDTs were mainly degraded in anaerobic condition in a typical tropical bay.
Our result is important for understanding distribution, source, and degradation of DDT in the typical tropical bay.
The utility of induced pluripotent stem cells (iPSCs) as models to study diseases and as sources for cell therapy depends on the integrity of their genomes. Despite recent publications of DNA ...sequence variations in the iPSCs, the true scope of such changes for the entire genome is not clear. Here we report the whole-genome sequencing of three human iPSC lines derived from two cell types of an adult donor by episomal vectors. The vector sequence was undetectable in the deeply sequenced iPSC lines. We identified 1,058–1,808 heterozygous single-nucleotide variants (SNVs), but no copy-number variants, in each iPSC line. Six to twelve of these SNVs were within coding regions in each iPSC line, but ∼50% of them are synonymous changes and the remaining are not selectively enriched for known genes associated with cancers. Our data thus suggest that episome-mediated reprogramming is not inherently mutagenic during integration-free iPSC induction.
► Deep whole-genome sequencing of three human iPSC lines generated with episomal vectors ► No vector sequence found in the nuclear and mitochondrial genomes ► Single-nucleotide and copy-number variation occurs at a normal frequency ► No evidence for selective enrichment of variation at functionally relevant loci
Some bladder-related diseases, such as bladder urinary tract infection (UTI) and bladder cancer (BCa), have significant six differences in incidence and prognosis. However, the molecular mechanisms ...underlying these sex differences are still not fully understood. Understanding the sex-biased differences in gene expression in normal bladder cells can help resolve these problems.
We first collected published single-cell RNA sequencing (scRNA-seq) data of normal human bladders from females and males to map the bladder transcriptomic landscape. Then, Gene Ontology (GO) analysis and gene set enrichment analysis (GSEA) were used to determine the significant pathways that changed in the specific cell populations. The Monocle2 package was performed to reconstruct the differentiation trajectories of fibroblasts. In addition, the scMetabolism package was used to analyze the metabolic activity at the single-cell level, and the SCENIC package was used to analyze the regulatory network.
In total, 27,437 cells passed stringent quality control, and eight main cell types in human bladder were identified according to classical markers. Sex-based differential gene expression profiles were mainly observed in human bladder urothelial cells, fibroblasts, B cells, and T cells. We found that urothelial cells in males demonstrated a higher growth rate. Moreover, female fibroblasts produced more extracellular matrix, including seven collagen genes that may mediate BCa progression. Furthermore, the results showed that B cells in female bladders exhibited more B-cell activated signals and a higher expression of immunoglobulin genes. We also found that T cells in female bladders exhibited more T-cell activated signals. These different biological functions and properties of these cell populations may correlate with sex differences in UTI and BCa, and result in different disease processes and outcomes.
Our study provides reasonable insights for further studies of sex-based physiological and pathological disparities in the human bladder, which will contribute to the understanding of epidemiological differences in UTI and BCa.
Various cells within the adrenal microenvironment are important in maintaining the body homeostasis. However, our understanding of adrenal disease pathogenesis is limited by an incomplete molecular ...characterization of the cell types responsible for the organ's multiple homeostatic functions. We report a cellular landscape of the human adrenal gland using single‐cell RNA sequencing. We reveal characteristic features of cell types within the human adrenal microenvironment and found immune activation of nonimmune cells in the adrenal endothelial cells. We also reveal that abundant immune cells occupied a lot of space in adrenal gland. Additionally, Sex‐related diversity in the adrenocortical cells and different gene expression profiles between the left and right adrenal gland are also observed at single‐cell resolution. Together, at single‐cell resolution, the transcriptomic map presents a comprehensive view of the human adrenal gland, which serves as a fundamental baseline description of this organ and paves a way for the further studies of adrenal diseases.
We report a cellular landscape of the human adrenal gland using single‐cell RNA sequencing. At single‐cell resolution, the transcriptomic map presents a comprehensive view of the human adrenal gland, which serves as a fundamental baseline description of this organ and paves a way for the further studies of adrenal diseases
A mammalian plasma membrane is a structure on which several layers of complexity are built. The first order of complexity comes from the heterogeneity of lipid-ordered domains. Gangliosides in ...concert with cholesterol are preferentially packed on the outer leaflet and form lipid-ordered domains, commonly known as lipid rafts. The formation and dynamics of these domains impact nearly all membrane protein functions and are an intensely studied topic. However, tools suited for lipid domain alteration are extremely limited. Currently, methyl-β-cyclodextrin (MβCD) appears to be the most common way to disrupt lipid domains, which is believed to operate
cholesterol extraction. This significantly limits our ability in membrane biophysics research. Previously, we found that
-(3-oxo-dodecanoyl) homoserine lactone (3oc), a small signaling chemical produced by
, is highly efficient in altering lipid-ordered domains. In this study, 3oc was compared with MβCD in a series of biochemical, biophysical, and cell biological analyses. Per molarity, 3oc is more efficient than MβCD in domain alteration and appears to better retain membrane lipids after treatment. This finding will provide an essential reagent in membrane biophysics research.
Background
The chemoresistance of prostate cancer (PCa) is invariably associated with the aggressiveness and metastasis of this disease. New emerging evidence indicates that the ...epithelial‐to‐mesenchymal transition (EMT) may play pivotal roles in the development of chemoresistance and metastasis. As a hallmark of EMT, E‐cadherin is suggested to be a key marker in the development of chemoresistance. However, the molecular mechanisms underlying PCa chemoresistance remain unclear. The current study aimed to explore the association between EMT and chemoresistance in PCa as well as whether changing the expression of E‐cadherin would affect PCa chemoresistance.
Methods
Parental PC3 and DU145 cells and their chemoresistant PC3‐TxR and DU145‐TxR cells were analyzed. PC3‐TxR and DU145‐TxR cells were transfected with E‐cadherin‐expressing lentivirus to overexpress E‐cadherin; PC3 and DU145 cells were transfected with small interfering RNA to silence E‐cadherin. Changes of EMT phenotype‐related markers and signaling pathways were assessed by Western blotting and quantitative real‐time polymerase chain reaction. Tumor cell migration, invasion, and colony formation were then evaluated by wound healing, transwell, and colony formation assays, respectively. The drug sensitivity was evaluated using MTS assay.
Results
Chemoresistant PC3‐TxR and DU145‐TxR cells exhibited an invasive and metastatic phenotype that associated with EMT, including the down‐regulation of E‐cadherin and up‐regulation of Vimentin, Snail, and N‐cadherin, comparing with that of parental PC3 and DU145 cells. When E‐cadherin was overexpressed in PC3‐TxR and DU145‐TxR cells, the expression of Vimentin and Claudin‐1 was down‐regulated, and tumor cell migration and invasion were inhibited. In particular, the sensitivity to paclitaxel was reactivated in E‐cadherin‐overexpressing PC3‐TxR and DU145‐TxR cells. When E‐cadherin expression was silenced in parental PC3 and DU145 cells, the expression of Vimentin and Snail was up‐regulated, and, particularly, the sensitivity to paclitaxel was decreased. Interestingly, Notch‐1 expression was up‐regulated in PC3‐TxR and DU145‐TxR cells, whereas the E‐cadherin expression was down‐regulated in these cells comparing with their parental cells. The use of γ‐secretase inhibitor, a Notch signaling pathway inhibitor, significantly increased the sensitivity of chemoresistant cells to paclitaxel.
Conclusion
The down‐regulation of E‐cadherin enhances PCa chemoresistance via Notch signaling, and inhibiting the Notch signaling pathway may reverse PCa chemoresistance.
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