Distinct genomic traits of acral and mucosal melanomas revealed by targeted mutational profiling

The incidence of melanoma is rising globally including China. Comparing to Caucasians, the incidence of non‐cutaneous melanomas is significantly higher in Chinese. Herein, we performed genomic profiling of 89 Chinese surgically resected primary melanomas, including acral (n = 54), cutaneous (n = 22), and mucosal (n = 13), by hybrid capture‐based next‐generation sequencing. We show that mucosal melanomas tended to harbor more pathogenic mutations than other types of melanoma, though the biological significance of this finding remains uncertain. Chromosomal arm‐level alterations including 6q, 9p, and 10p/q loss were highly recurrent in all subtypes, but mucosal melanoma was significantly associated with increased genomic instability. Importantly, 7p gain significantly correlated with unfavorable clinical outcomes in non‐cutaneous melanomas, representing an intriguing prognostic biomarker of those subtypes. Furthermore, focal amplification of 4q12 (KIT, KDR, and PDGFRα) and RAD51 deletion were more abundant in mucosal melanoma, while NOTCH2 amplification was enriched in acral melanoma. Additionally, cutaneous melanomas had higher mutation load than acral melanomas, while mucosal melanomas did not differ from other subtypes in mutation burden. Together, our data revealed important features of acral and mucosal melanomas in Chinese including distinctive driver mutation pattern and increased genomic instability. These findings highlight the possibilities of combination therapies in the clinical management of melanoma.     

Advancing Aptamers as Molecular Probes for Cancer Theranostic Applications—The Role of Molecular Dynamics Simulation

Theranostics cover emerging technologies for cell biomarking for disease diagnosis and targeted introduction of drug ingredients to specific malignant sites. Theranostics development has become a significant biomedical research endeavor for effective diagnosis and treatment of diseases, especially cancer. An efficient biomarking and targeted delivery strategy for theranostic applications requires effective molecular coupling of binding ligands with high affinities to specific receptors on the cancer cell surface. Bioaffinity offers a unique mechanism to bind specific target and receptor molecules from a range of non‐targets. The binding efficacy depends on the specificity of the affinity ligand toward the target molecule even at low concentrations. Aptamers are fragments of genetic materials, peptides, or oligonucleotides which possess enhanced specificity in targeting desired cell surface receptor molecules. Aptamer–target binding results from several inter‐molecular interactions including hydrogen bond formation, aromatic stacking of flat moieties, hydrophobic interaction, electrostatic, and van der Waals interactions. Advancements in Systematic Evolution of Ligands by Exponential Enrichment (SELEX) assay has created the opportunity to artificially generate aptamers that specifically bind to desired cancer and tumor surface receptors with high affinities. This article discusses the potential application of molecular dynamics (MD) simulation to advance aptamer‐mediated receptor targeting in targeted cancer therapy. MD simulation offers real‐time analysis of the molecular drivers of the aptamer‐receptor binding and generate optimal receptor binding conditions for theranostic applications. The article also provides an overview of different cancer types with focus on receptor biomarking and targeted treatment approaches, conventional molecular probes, and aptamers that have been explored for cancer cells targeting.     

Overexpression of ER Protein Processing and Apoptosis Regulator Genes in Human Embryonic Kidney 293 Cells Improves Gene Therapy Vectors Production

Gammaretroviral and lentiviral vectors (γ‐RV and LV) are among the most used vectors in gene therapy. Currently, human embryonic kidney (HEK) 293 cells, the manufacture platform of choice for these vectors, provide low transducing particle yields, challenging their therapeutic applications and commercialization. This work studies metabolic pathways, focusing on endoplasmic reticulum (ER) protein processing and anti‐apoptotic mechanisms, influencing vector productivity in HEK 293 cell substrates. To that end, four candidate genes—protein disulfide isomerase family A member 2 gene, heat shock protein family A (Hsp70) member 5 gene, X‐box binding protein 1 gene (ER protein processing), and B‐cell lymphoma 2 protein gene (anti‐apoptotic)—are individually stably expressed in the cells. How their overexpression level influence vector yields is analyzed by establishing cell populations with incremental genomic copies of each. γ‐RV volumetric productivity increases up to 97% when overexpressing ER protein processing genes. LV volumetric production increases 53% when overexpressing the anti‐apoptotic gene. Improvements are associated with higher cell specific productivities and dependent on gene overexpression level, highlighting the importance of fine‐tuning gene expression. Overall, this work discloses gene engineering targets enabling efficient gene therapy product manufacture showing that ER protein processing and anti‐apoptotic pathways are pivotal to producer cell performance.     

Design and synthesis of a folate-receptor targeted diazepine-ring-opened pyrrolobenzodiazepine prodrug conjugate

Pyrrolobenzodiazepines (PBDs) and their dimers (bis-PBDs) have emerged as some of the most potent chemotherapeutic compounds and are currently under development as novel payloads in antibody-drug conjugates (ADCs). However, when used as stand-alone therapeutics or as warheads for small molecule drug conjugates (SMDCs), dose-limiting toxicities are often observed. As an elegant solution to this inherent problem, we designed and synthesized a diazepine-ring-opened bis-PBD prodrug (pro-PBD-PBD) folate conjugate lacking the one of the two imine moieties found in the corresponding free bis-PBD. Upon entering a targeted cell, cleavage of the linker system, including the hydrolysis of an oxazolidine moiety, results in the formation of a reactive intermediate which possesses a newly formed aldehyde as well as an aromatic amine. A fast and spontaneous intramolecular ring-closing reaction subsequently takes place as the aromatic amine adds to the aldehyde with the loss of water to give the imine, and as a result, the diazepine ring, thereby delivering the bis-PBD to the targeted cell. The in vitro and in vivo activity of this conjugate has been evaluated on folate receptor positive KB cells. Sub-nanomolar activity with good specificity and high cure rates with minimal toxicity have been observed.     

Improving NK1R-targeted gene delivery of stearyl-antimicrobial peptide CAMEL by conjugating it with substance P

Specificity is a crucial condition that hampers the application of non-viral vectors for cancer gene therapy. In a previous study, we developed an efficient gene vector, stearyl-CAMEL, using N-terminal stearylation of the antimicrobial peptide CAMEL. Substance P (SP), an 11-residue neuropeptide, rapidly enters cells after binding to the neurokinin-1 receptor (NK1R), which is expressed in many cancer cell lines. In this study, the NK1R-targeted gene vector stearyl-CMSP was constructed by conjugating SP to the C-terminus of stearyl-CAMEL. Our results indicated that stearyl-CMSP displayed significant transfection specificity for NK1R-expressing cells compared with that shown by stearyl-CAMEL. Accordingly, the stearyl-CMSP/p53 plasmid complexes had significantly higher antiproliferative activity against HEK293-NK1R cells than they did against HEK293 cells, while the stearyl-CAMEL/p53 plasmid complexes did not show this specificity in antiproliferative activity. Consequently, conjugation of the NK1R-targeted ligand SP is a simple and successful strategy to construct efficient cancer-targeted non-viral gene vectors.     

干扰素阴道胶囊联合微波治疗宫颈炎合并高危型HPV感染对外周血Th17和Treg细胞及炎症因子的影响

目的探讨干扰素阴道胶囊联合微波治疗宫颈炎合并高危型人乳头瘤病毒(HPV)感染对外周血Th17和Treg细胞及炎症因子的影响。方法随机选择2015年1月-2018年1月在本院就诊的慢性宫颈炎合并高危型HPV感染患者90例,按随机数字表法分为观察组和对照组,每组45例。对照组给予重组人干扰素α2b栓,观察组给予重组人干扰素α2b栓联合微波治疗,治疗3个月后,观察两组患者的症状改善情况、HPV转阴率、血清炎性因子水平以及Th17和Treg细胞水平。结果观察组总有效为41例,占91.1%,对照组为73.3%,观察组显著高于对照组(P<0.05)。治疗后观察组的白带量改善率、白带脓性改善率、高危型HPV转阴率均显著高于对照组(P<0.05)。治疗前两组患者的白介素17(IL17)、肿瘤坏死因子β(TNFβ)、白介素23(IL23)水平比较,差异无统计学意义(P>0.05);治疗后两组患者的IL17、TNFβ、IL23水平均显著下降,且观察组下降更加显著(P<0.05)。治疗后观察组的Treg细胞、Th17细胞水平,Th17/Treg比值均较对照组低(P<0.05)。治疗前两组患者的滴虫、真菌、细菌性阴道炎、衣原体、解脲支原体感染人数比较差异无统计学意义(P>0.05),治疗后观察组的细菌性阴道炎、衣原体、解脲支原体感染显著降低,与对照组比较差异有统计学意义(P<0.05)。结论干扰素阴道胶囊联合微波治疗宫颈炎患者的治疗效果好,可显著改善患者症状,减轻体内炎症反应,增加HPV转阴率,调节Th17和Treg细胞,对预防宫颈癌有重要作用。     

精准营养联合生长抑素对胃癌根治术患者的临床疗效

目的探讨生长抑素联合精准营养疗法对胃癌根治术患者术后营养指标水平和肠道菌群的影响。方法选取我院2017年10月至2019年1月收治并拟行胃癌根治术治疗的92例胃癌患者为研究对象,随机分为对照组(46例)和观察组(46例)。两组患者术后均给予常规处理。对照组患者同时给予生长抑素+常规肠外营养支持,观察组患者同时给予生长抑素+精准营养支持。于营养干预前后统计两组患者营养相关指标[体质量、血清白蛋白(ALB)、前白蛋白(PA)、淋巴细胞计数(LC)、肌酐(Ct)以及尿素氮(UN)]水平;对肠道菌群进行alpha分析,并计算肠杆菌、肠球菌、乳杆菌和双歧杆菌数量。结果营养干预前,两组患者营养相关指标水平以及肠道菌群相关指标水平之间差异无统计学意义(均P>0.05)。营养干预后,(1)观察组患者平均体质量以及血清ALB、PA、LC水平分别为(56.98±5.34)kg、(29.98±6.03)mg/L、(137.14±35.17)g/L、(1.15±0.26)×10⁹,对照组分别为(53.29±5.18)kg、(26.12±5.29)mg/L、(104.53±27.66)g/L、(0.82±0.18)×10⁹,两组之间差异有统计学意义(均P<0.05);(2)观察组患者肠道菌群Chao指数和Ace指数水平分别为(397.84±75.23)、(408.26±78.34),对照组分别为(362.13±66.23)、(371.19±70.09),两组之间差异有统计学意义(均P<0.05);(3)观察组患者肠道肠杆菌、肠球菌数量明显低于对照组,而乳杆菌、双歧杆菌数量明显高于对照组(均P<0.05)。结论胃癌根治术患者术后联合生长抑素和精准营养疗法能够显著改善患者的营养状况,保持肠道菌群丰度,平衡肠道菌群分布,值得临床研究和应用。     

Efficient Editing of an Adenoviral Vector Genome with CRISPR/Cas9

Immunotherapy based on genetic modification of T cells has played an important role in the treatment of tumors and viral infections. Moreover, adenoviral vectors engineered with improved safety due to their inability to integrate into the host genome have been key in the clinical application of T cell therapy. However, the commonly used adenoviral vector Ad5 exhibits low efficiency of infection of human T cells and the details of the intracellular trafficking pathway of adenoviral vectors in human primary T cells remains unclear. Resolution of these issues will depend on successful modification of the adenoviral vector. To this end, here we describe the successful establishment of a simple and efficient method for editing adenoviral vectors in vitro using the CRISPR-Cas9 gene editing system to target the adenoviral fiber gene. Electronic supplementary materialThe online version of this article (10.1007/s12088-020-00905-3) contains supplementary material, which is available to authorized users.     

Efecto positivo de la terapia de luz brillante sobre el estado de ánimo y la calidad del sueño en las personas mayores institucionalizadas

IntroductionBright light exposure during the day has a positive effect on health and its deficit can cause multiple physiological and cognitive disorders, including depression. The aim of this study was to evaluate the effect of bright light therapy (BLT) on the quality of sleep and mood emotional state; cognitive status, global deterioration and quality of life in institutionalized elderly.Material and methodsThis is a study with repeated measures design. Thirty-seven older people admitted to a nursing home. The study lasted 3 weeks. The first week, the reference values were established with the Oviedo Sleep Questionnaire, Yesavage Depression Scale, Mini-Mental, Global Scale of Impairment and European Quality of Life Questionnaire. During the second week, they were exposed to BLT (7,000-10,000 lx at eye level) between 9:30 a.m. and 11:00 a.m. During the third week, all the data were re-evaluated.ResultsAll variables improved significantly after the application of light therapy. Sleep (COS) pre-test 4.1 ± 1.49, post-test 4.9 ± 1.46, p: 0.01), mood (pre-test 3.65 ± 2.78, post-test 2.65 ± 2.9, p: 0.01), cognitive state (pre-test 22.72 ± 6.53, post-test 24 ± 5.92, p: 0.001), state of global deterioration (pre-test 3.10 ± 1.26, post-test 2.72 ± 5.92, p: 0.001) and health-related quality of life (pre-test 6.93 ± 1.86, post-test 7.82 ± 1.62, p: 0.001).ConclusionsSleep quality, mood, cognitive status, global deterioration status and quality of life significantly improved after the application of light bright therapy.