Patient-derived organoids (PDO) of lung cancer has been recently introduced, reflecting the genomic landscape of lung cancer. However, clinical relevance of advanced lung adenocarcinoma organoids ...remains unknown. Here, we examined the ability of PDOs to predict clinical responses to targeted therapies in individual patients and to identify effective anticancer therapies for novel molecular targets.
Eighty-four organoids were established from patients with advanced lung adenocarcinoma. Formalin-fixed, paraffin-embedded tumor specimens from corresponding patients were analyzed by whole-exome sequencing (
= 12). Organoids were analyzed by whole-exome sequencing (
= 61) and RNA sequencing (
= 55). Responses to mono or combination targeted therapies were examined in organoids and organoid-derived xenografts.
PDOs largely retained somatic alterations including driver mutations of matching patient tumors. PDOs were able to recapitulate progression-free survival and objective responses of patients with non-small cell lung cancer receiving clinically approved tyrosine kinase inhibitors. PDOs recapitulated activity of therapeutic strategies under clinical investigation. YUO-071 harboring an
exon 19 deletion and a
G464A mutation and the matching patient responded to dabrafenib/trametinib combination therapy. YUO-004 and YUO-050 harboring an
L747P mutation was sensitive to afatinib, consistent with the response in the matching patient of YUO-050. Furthermore, we utilized organoids to identify effective therapies for novel molecular targets by demonstrating the efficacy of poziotinib against
exon 20 insertions and pralsetinib against
fusions.
We demonstrated translational relevance of PDOs in advanced lung adenocarcinoma. PDOs are an important diagnostic tool, which can assist clinical decision making and accelerate development of therapeutic strategies.
Plants produce and accumulate stress-resistant substances when exposed to abiotic stress, which involves a protein conversion mechanism that breaks down stress-damaged proteins and supplies usable ...amino acids. Eukaryotic protein turnover is mostly driven by the ubiquitination pathway. Among the three enzymes required for protein degradation, E3 ubiquitin ligase plays a pivotal role in most cells, as it determines the specificity of ubiquitination and selects target proteins for degradation. In this study, to investigate the function of
(Plant U-box gene in
), we constructed a CRISPR/Cas9 vector, generated
gene-edited individuals, and evaluated resistance to abiotic stress using gene-edited lines. A stress-tolerant phenotype was observed as a result of drought and salinity stress treatment in the T
gene-edited null lines (PUB7-GE) lacking the T-DNA. In addition, although PUB7-GE did not show any significant change in mRNA expression analysis, it showed lower ion leakage and higher proline content than the wild type (WT). Protein-protein interaction analysis revealed that the expression of the genes (
,
,
, and
) known to be involved in stress increased in PUB7-GE and this, by forming a 1-node network with
and
, acted as a negative regulator of drought and salinity stress. This result provides evidence that
will be a useful target for both breeding and future research on drought tolerance/abiotic stress in rice.
Left ventricular hypertrophy (LVH) and left ventricular diastolic dysfunction (LVDD) are highly prevalent predictors of cardiovascular disease in individuals with chronic kidney disease (CKD). ...Vitamin D, particularly 25-hydroxyvitamin D 25(OH)D, deficiency has been reported to be associated with cardiac structure and function in CKD patients. In the current study, we investigated the association between 1,25-dihydroxyvitamin D 1,25(OH)2D, the active form of 25(OH)D, and LVH/LVDD in CKD patients. We enrolled 513 non-dialysis CKD patients. The presence of LVH and LVDD was determined using transthoracic echocardiography. In multivariable analysis, serum 1,25(OH)2D levels, but not serum 25(OH)D, were independently associated with LVH odds ratio (OR): 0.90, 95% confidential interval (CI): 0.88-0.93, P < 0.001. Additionally, age, systolic blood pressure, and intact parathyroid hormone levels were independently associated with LVH. Similarly, multivariable analysis demonstrated that serum 1,25(OH)2D levels, but not 25(OH)D levels, were independently associated with LVDD (OR: 0.88, 95% CI: 0.86-0.91, P < 0.001) with systolic blood pressure showing independent association with LVDD. The optimal cut-off values for serum 1,25(OH)2D levels for identifying LVH and LVDD were determined as ≤ 12.7 pg/dl and ≤ 18.1 pg/dl, respectively. Our findings suggest that serum 1,25(OH)2D levels have independent association with LVH and LVDD in CKD patients, underscoring their potential as biomarkers for these conditions in this patient population.
Cardiac valve calcification is highly prevalent in patients with chronic kidney disease (CKD). Low vitamin D levels are associated with vascular calcification in CKD. However, the association between ...vitamin D levels and cardiac valve calcification is unknown. A total of 513 patients with pre-dialysis CKD were included in this cross-sectional study. Aortic valve calcification (AVC) and mitral valve calcification (MVC) were assessed using two-dimensional echocardiography. The associations between AVC and MVC and baseline variables were investigated using logistic regression analyses. In multivariable analysis, serum 1,25(OH)
D level was independently associated with AVC (odds ratio OR, 0.87; P < 0.001) and MVC (OR, 0.92; P < 0.001). Additionally, age, diabetes, coronary heart disease, calcium × phosphate product, and intact parathyroid hormone levels were independently associated with AVC and MVC. Systolic blood pressure was independently associated with AVC. A receiver-operating characteristic (ROC) curve analysis showed that the best cutoff values of serum 1,25(OH)
D levels for predicting AVC and MVC were ≤ 12.5 and ≤ 11.9 pg/dl, respectively. Serum 1,25(OH)
D deficiency is independently associated with AVC and MVC in patients with CKD, suggesting that serum 1,25(OH)
D level may be a potential biomarker of AVC and MVC in these patients.
Procalcitonin (PCT) is a biomarker for diagnosing infections and guiding antibiotic therapy. In this study, we investigated whether PCT can predict survival and recovery at 28 days in critically ill ...patients with sepsis-induced acute kidney injury (SIAKI) receiving continuous renal replacement therapy (CRRT). We examined 649 patients with SIAKI who underwent CRRT in our intensive care unit. In a multivariable Cox regression analysis, a single PCT level at CRRT initiation was not associated with survival in all patients. However, the higher % PCT decrease over 72 hours after CRRT initiation was independently associated with the higher chance of 28-day survival (per 10% decrease, hazard ratio HR for mortality: 0.87, 95% confidence interval CI: 0.85-0.89; P < 0.001). Among the survivors, the % PCT decrease over 72 hours after CRRT initiation, not a single PCT level at CRRT initiation, was independently associated with recovery from dialysis (per 10% decrease, HR for renal recovery: 1.28, 95% CI:1.21-1.36; P < 0.001). This study demonstrated that the higher % PCT decrease was independently associated with the higher chance of survival and recovery from dialysis at 28 days in critically ill patients with SIAKI receiving CRRT. Thus, a decrease in the PCT level, not a single PCT level at CRRT initiation, could be a valuable tool for predicting prognosis in these patients.
Infectious diseases such as Coronavirus disease 2019 (COVID‐19) and Middle East respiratory syndrome (MERS) present an increasingly persistent crisis in many parts of the world. COVID‐19 is caused by ...severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). The angiotensin‐converting enzyme 2 (ACE2) is a crucial cellular receptor for SARS‐CoV‐2 infection. Inhibition of the interaction between SARS‐CoV‐2 and ACE2 has been proposed as a target for the prevention and treatment of COVID‐19. We produced four recombinant plant‐derived ACE2 isoforms with or without the mu tailpiece (μ‐tp) of immunoglobulin M (IgM) and the KDEL endoplasmic reticulum retention motif in a plant expression system. The plant‐derived ACE2 isoforms bound whole SARS‐CoV‐2 virus and the isolated receptor binding domains of SARS‐CoV‐2 Alpha, Beta, Gamma, Delta, and Omicron variants. Fusion of μ‐tp and KDEL to the ACE2 protein (ACE2 μK) had enhanced binding activity with SARS‐CoV‐2 in comparison with unmodified ACE2 protein derived from CHO cells. Furthermore, the plant‐derived ACE2 μK protein exhibited no cytotoxic effects on Vero E6 cells and effectively inhibited SARS‐CoV‐2 infection. The efficient and rapid scalability of plant‐derived ACE2 μK protein offers potential for the development of preventive and therapeutic agents in the early response to future viral outbreaks.
Graphical and Lay Summary
In this study, we focused on the efficient, rapid, and scalable expression of ACE2 protein as a preventive and therapeutic agent for the future outbreaks of SARS‐CoV‐2 and other viral pandemics using Nicotiana benthamiana as a plant transient expression system. To enhance the functionality of the protein, we endeavored to produce oligomeric ACE2 µK by incorporating the KDEL endoplasmic reticulum retention motif and mu tailpiece (μ‐tp) of immunoglobulin M, while excluding the Fc region of the antibody. The plant‐derived ACE2 μK protein exhibited enhanced binding activity against various SARS‐CoV‐2 strains in comparison to ACE2 protein derived from CHO cells. Moreover, the ACE2 μK protein exhibited no cytotoxic effects on Vero E6 cells and effectively inhibited SARS‐CoV‐2 infection. To our knowledge, this work is the first attempt to fuse the μ‐tp without the Fc region to enhance the activity of a recombinant protein. The graphical was created with BioRender.com.
Cell stress may give rise to insuperable growth arrest, which is defined as cellular senescence. Stenotic kidney (STK) ischemia and injury induced by renal artery stenosis (RAS) may be associated ...with cellular senescence. Mesenchymal stem cells (MSCs) decrease some forms of STK injury, but their ability to reverse senescence in RAS remains unknown. We hypothesized that RAS evokes STK senescence, which would be ameliorated by MSCs. Mice were studied after 4 weeks of RAS, RAS treated with adipose tissue‐derived MSCs 2 weeks earlier, or sham. STK senescence‐associated β‐galactosidase (SA‐β‐Gal) activity was measured. Protein and gene expression was used to assess senescence and the senescence‐associated secretory phenotype (SASP), and staining for renal fibrosis, inflammation, and capillary density. In addition, senescence was assessed as p16+ and p21+ urinary exosomes in patients with renovascular hypertension (RVH) without or 3 months after autologous adipose tissue‐derived MSC delivery, and in healthy volunteers (HV). In RAS mice, STK SA‐β‐Gal activity increased, and senescence and SASP marker expression was markedly elevated. MSCs improved renal function, fibrosis, inflammation, and capillary density, and attenuated SA‐β‐Gal activity, but most senescence and SASP levels remained unchanged. Congruently, in human RVH, p21+ urinary exosomes were elevated compared to HV, and only slightly improved by MSC, whereas p16+ exosomes remained unchanged. Therefore, RAS triggers renal senescence in both mice and human subjects. MSCs decrease renal injury, but only partly mitigate renal senescence. These observations support exploration of targeted senolytic therapy in RAS.
Stenotic kidney (STK) ischemia induced by renal artery stenosis (RAS) is associated with cellular senescence in mice and human subjects. Mesenchymal stem cell (MSC) treatment decreases some forms of STK injury, but only partly mitigates renal senescence.
Tooth-bone-borne rapid palatal expansion (RPE) often results in unwanted buccal tipping of anchor teeth, particularly in cases where midpalatal suture expansion is not successful. This report ...presents a novel approach to treating transverse maxillary deficiency through purely bone-borne RPE using bracket plates. The method involves strategically placing four mini-implants in the palate before inserting the RPE screw. Subsequently, right and left bracket plates are fixed over the mini-implant heads to connect two anteroposterior mini-implants. The RPE screw is then attached to the bracket plates, securing the RPE connecting arms into 0.032 × 0.040-inch slot brackets on the bracket plates. The primary advantage of this method is that the RPE screw is replaceable and adjustable. This enhances the freedom in selecting mini-implant placement sites, ensuring better skeletal anchorage on the palatal bone compared to tooth-bone-borne RPE, where mini-implant placement sites are limited to the periphery of the RPE screw. This report presents two cases in which midpalatal suture expansion was successfully achieved using purely bone-borne RPE without incurring any dental side effects.
Although nodal metastasis (NM) is an important prognostic factor of ampullary adenocarcinoma, the prognostic implication of extranodal extension (ENE) is not well characterized. NM with ENE status ...was investigated in 279 surgically resected ampullary adenocarcinoma patients and compared with other clinicopathologic factors, including overall survival (OS) and recurrence-free survival (RFS). Expression of epithelial–mesenchymal transition (EMT) markers, including E-cadherin, Twist, and Snail, was assessed in a subset of the cohort. NM was observed in 94 cases (33.7%), of which ENE was observed in 32 cases (34%). NM with ENE was more frequently associated with tumors with poor differentiation than NM without ENE (P = .017). The 5-year OS and RFS rates of patients with NM and ENE was significantly worse (13.0% and 6.3%) than those with NM without ENE (37.7% and 21.4%) and those without NM (57.6% and 50.2%, respectively; P < .001). When pN category was matched, the OS and RFS was worse in patients with ENE than in those without ENE (P < .05). Moreover, the expression of E-cadherin and Twist was significantly different between NM areas with and without ENE (all, P < .001). Since ENE was associated with poorly differentiated ampullary adenocarcinomas and showed different expression of EMT markers, EMT could be a possible mechanism of ENE. Ampullary adenocarcinoma patients with ENE had worse OS and RFS than those without ENE. Therefore, evaluation of ENE can provide additional survival information for patients with surgically resected ampullary carcinoma.
•Ampullary adenocarcinoma patients with extranodal extension (ENE) showed worse survival rate than those without ENE.•Epithelial-mesenchymal transition could be a possible mechanism of ENE.
Ischemia-reperfusion injury (IRI) in the kidneys is a major cause of acute kidney injury (AKI). Time-restricted feeding (TRF), known for its metabolic health benefits and alleviation of various ...chronic diseases without calorie restriction, was investigated for its potential protective effects against IRI-induced AKI. Male C57BL/6 mice underwent unilateral IRI, with their kidneys collected after two days. For two weeks before IRI induction, the TRF group had unlimited access to standard chow but within an 8-hour feeding window during the dark cycle. The study groups were Control, TRF, IRI, and TRF + IRI. In the TRF + IRI group, tubular damage scores significantly decreased compared to the IRI group. Furthermore, the TRF + IRI mice had lower levels of phosphorylated NF-κB and fewer F4/80-positive macrophages than the IRI group. Oxidative stress markers for lipids and proteins were also notably lower in the TRF + IRI group. Additionally, TUNEL-positive tubular cells and cleaved caspase-3 expression were reduced in the TRF + IRI group. Without calorie restriction, TRF mitigated renal damage by reducing inflammation, oxidative stress, and tubular apoptosis in renal IRI. This suggests that TRF could be a promising dietary strategy to prevent IRI-induced AKI.
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