This study aimed to evaluate the usefulness of four microRNAs (miRNAs) in an acute pancreatic injury dog model. Acute pancreatitis was induced by infusion of cerulein for 2 h (7.5 μg/kg/h). The levels of well-known miRNAs, microRNA-216a (miR-216a) and microRNA-375 (miR-375), and new candidates microRNA-551b (miR-551b), and microRNA-7 (miR-7), were measured at 0, 0.5, 1, 2, 6, 12, and 24 h with serum amylase and lipase, and histopathological examination was performed. Among the four miRNAs, miR-216a and miR-375, and serum enzymes were significantly increased by cerulein treatment. The expression levels of miRNAs and serum enzymes peaked at 2–6 h with a similar pattern; however, the overall increases in miR-216a and miR-375 levels were much higher than those of the serum enzyme biomarkers. Increased levels of miR-216a and miR-375 were most highly correlated to the degree of individual histopathological injuries of the pancreas, and showed much greater dynamic response than serum enzyme biomarkers. Twenty-four-hour time-course analysis in this study revealed time-dependent changes of miRNA expression levels, from initial increase to decrease by predose level in acute pancreatitis. Our findings demonstrate that, in dogs, miR-216a and miR-375 have the potential to sensitively detect pancreatitis and reflect well the degree of pancreatic injury, whereas miR-551b and miR-7 do not.

Drug-induced phospholipidosis (PL) is a storage disorder caused by the formation of phospholipid-drug complexes in lysosomes. Because of the diversity of PL between species, human cell-based assays have been used to predict drug-induced PL in humans. We established three-dimensional (3D) human liver organoids as described previously and investigated their liver characteristics through multiple analyses. Drug-induced PL was initiated in these organoids and in monolayer HepG2 cultures, and cellular changes were systemically examined. Organoids that underwent differentiation showed characteristics of hepatocytes rather than HepG2 cells. The organoids also survived under PL-inducing drug conditions for 48 h and maintained a more stable albumin secretion level than the HepG2 cells. More cytoplasmic vacuoles were observed in organoids and HepG2 cells treated with more potent PL-induced drugs, but to a greater extent in organoids than in HepG2 cells. Lysosome-associated membrane protein 2, a marker of lysosome membranes, showed a stronger immunohistochemical signal in the organoids. PL-distinctive lamellar bodies were observed only in amiodarone-treated organoids by transmission electron microscopy. Human liver organoids are thus more sensitive to drug-induced PL and less affected by cytotoxicity than HepG2 cells. Since PL is a chronic condition, these results indicate that organoids better reflect metabolite-mediated hepatotoxicity in vivo and could be a valuable system for evaluating the phospholipidogenic effects of different compounds during drug development.

Tumor innervation has been shown to be utilized by some solid cancers to support tumor initiation, growth, progression, and metastasis, as well as confer resistance to immune checkpoint blockade through suppression of antitumor immunologic responses. Since botulinum neurotoxin type A1 (BoNT/A1) blocks neuronal cholinergic signaling, its potential use as an anticancer drug in combination with anti-PD-1 therapy was investigated in four different syngeneic mouse tumor models. Mice implanted with breast (4T1), lung (LLC1), colon (MC38), and melanoma (B16-F10) tumors were administered a single intratumoral injection of 15 U/kg BoNT/A1, repeated intraperitoneal injections of 5 mg/kg anti-PD-1 (RMP1-14), or both. Compared to the single-agent treatments, anti-PD-1 and BoNT/A1 combination treatment elicited significant reduction in tumor growth among B16-F10 and MC38 tumor-bearing mice. The combination treatment also lowered serum exosome levels in these mice compared to the placebo control group. In the B16-F10 syngeneic mouse tumor model, anti-PD-1 + BoNT/A1 combination treatment lowered the proportion of MDSCs, negated the increased proportion of T_reg cells, and elicited a higher number of tumor-infiltrating CD4⁺ and CD8⁺ T lymphocytes into the tumor microenvironment compared to anti-PD-1 treatment alone. Our findings demonstrate the synergistic antitumor effects of BoNT/A1 and PD-1 checkpoint blockade in mouse tumor models of melanoma and colon carcinoma. These findings provide some evidence on the potential application of BoNT/A1 as an anticancer drug in combination with immune checkpoint blockade and should be further explored.

Phthalate esters (PEs) are the most widely used class of plasticizers. Several PEs, however, were found to have adverse effects on the health of animals. A new phthalate-free plasticizer, Eco-DEHCH (bis(2-ethylhexyl) cyclohexane-1,4-dicarboxylate), was recently developed as an ecofriendly replacement for phthalate plasticizers and to be less harmful to organisms. The present study evaluated the long-term toxicity of Eco-DEHCH in Wistar Han rats to explore adverse effects and predict hazardous potential to humans. Forty male and forty female Wistar Han rats were exposed to Eco-DEHCH in dietary feed for 52 weeks, and their hematologic, coagulation, and serum biochemical parameters were monitored. The rats were subjected to close clinical, ophthalmic, and histopathologic examinations and urinalysis throughout the consumption of Eco-DEHCH. The effects of this plasticizer on food consumption and organ weight were also determined. Chronic exposure to Eco-DEHCH was generally safe, although it also resulted in α_2u-globulin accumulation, a parameter with no human relevance. In conclusion, Eco-DEHCH can serve as a safe and promising alternative plasticizer.

L1 cell adhesion molecule (L1CAM, CD171) is a transmembrane glycoprotein important for nervous system development. Conversely, L1CAM is overexpressed in many human solid tumors with poor prognosis. Nonclinical safety evaluation and pharmacokinetics (PK) of Ab612, a human anti-L1CAM monoclonal antibody (mAb), were conducted to support a first-in-human (FIH) clinical trial for the treatment of L1CAM overexpressing tumors. A tissue cross-reactivity (TCR) assessment showed that Ab612 exhibited staining in various tissues from mouse, rat, canine, cynomolgus monkey, and human, including those associated with peripheral nerves, myenteric plexus, spinal nerve roots, and certain cell types, such as mononuclear cells (lymphocytes), cardiac Purkinje fibers, and epithelial cell in the lung and kidney. These findings were consistent with the reported expression of the L1CAM by these cell types. Dose range-finding (DRF) studies were conducted in rats and cynomolgus monkeys at doses up to 500 and 400 mg/kg, respectively, by intravenous (IV) administration once weekly for 2 weeks. Results showed no treatment-related adverse findings, and the maximum tolerated dose (MTD) was the highest dose tested. Four-week repeat-dose toxicity studies were conducted in rats and cynomolgus monkeys at doses up to MTD, by IV administration once weekly for 4 weeks with a 28-day recovery period. Results showed no treatment-related adverse findings, and the no-observed-adverse-effect-level (NOAEL) was the highest dose tested. Single dose PK studies in rats and cynomolgus monkeys with doses of 5–100 and 5–50 mg/kg, respectively, confirmed the PK in the expected range for mAbs. These preclinical data indicate no safety concerns and provide adequate safety margins for the planned dose levels in the FIH trial.

유방암은 다양한 생물학적 요소로 구성된 복잡한 질병으로, 여러 분자적 서브 타입을 유발한다. 정확한 서브 타입 예측은 암의 예후에 중대한 영향을 가지며, 서브 타입별 치료법 제공을 통한 환자의 생존율 향상에 중요하나, 생물학적 이질성으로 인해 쉽지 않다. 최근, 유전체 및 후성 유전체 데이터를 처리하기 위해 머신러닝 모델들이 유방암 분류에 적용되었으며, 특히 다중 오믹스를 활용한 연구들이 제시되었다. 하지만, 높은 차원과 복잡성으로 인해 특징 분석 및 분류 정확성에 한계를 갖는다. 본 논문에서는 뉴럴 네트워크를 기반으로 다중 오믹스 통합 데이터를 활용한 유방암 서브 타입 분류 모델을 제시한다. 유전자 발현, DNA 메틸레이션, 그리고 miRNA 오믹스를 통합한 데이터로 분류 모델을 구축하였으며, 성능 비교 결과, 평균 90.45%의 정확도로 기존 연구보다 높은 성능을 보였다. 제안된 모델을 통해 정확한 유방암 환자의 서브 타입 예측을 기반으로 환자의 예후 향상에 도움을 줄 것으로 기대된다.