In-Depth Structure and Function Characterization of Heterogeneous Interchain Cysteine-Conjugated Antibody-Drug Conjugates.

Antibody-drug conjugates (ADCs), integrating high specificity of antigen-targeting antibodies and high potency of cell-killing chemical drugs, have become one of the most rapidly expanding therapeutic biologics in oncology. Although ADCs were widely studied from multiple aspects, overall structural elucidation with comprehensive understanding of variants is scarcely reported. Here, for the first time, we present a holistic and in-depth characterization of an interchain cysteine-conjugated ADC, focusing on conjugation and charge heterogeneity, and in vitro biological activities. Conjugation mapping utilized a bottom-up approach, unraveled positional isomer composition, provided insights into the conjugation process, and elucidated how conjugation affects the physicochemical and biological properties of an ADC. Charge profiling combined bottom-up and top-down approaches to interrogate the origin of charge heterogeneity, its impact on function, and best practice for characterization. Specifically, we pioneered the utilization of capillary isoelectric focusing-mass spectrometry to decode not only critical post-translational modifications but also drug load and positional isomer distribution. The study design provides general guidance for in-depth characterization of ADCs, and the analytical findings in turn benefit the discovery and development of future ADCs.

An intrinsic tumour eviction mechanism in Drosophila mediated by steroid hormone signalling.

Polycomb group proteins are epigenetic regulators maintaining transcriptional memory during cellular proliferation. In Drosophila larvae, malfunction of Polyhomeotic (Ph), a member of the PRC1 silencing complex, results in neoplastic growth. Here, we report an intrinsic tumour suppression mechanism mediated by the steroid hormone ecdysone during metamorphosis. Ecdysone alters neoplastic growth into a nontumorigenic state of the mutant ph cells which then become eliminated during adult stage. We demonstrate that ecdysone exerts this function by inducing a heterochronic network encompassing the activation of the microRNA lethal-7, which suppresses its target gene chronologically inappropriate morphogenesis. This pathway can also promote remission of brain tumours formed in brain tumour mutants, revealing a restraining of neoplastic growth in different tumour types. Given the conserved role of let-7, the identification and molecular characterization of this innate tumour eviction mechanism in flies might provide important clues towards the exploitation of related pathways for human tumour therapy.

A switch in transcription and cell fate governs the onset of an epigenetically-deregulated tumor in Drosophila.

Tumor initiation is often linked to a loss of cellular identity. Transcriptional programs determining cellular identity are preserved by epigenetically-acting chromatin factors. Although such regulators are among the most frequently mutated genes in cancer, it is not well understood how an abnormal epigenetic condition contributes to tumor onset. In this work, we investigated the gene signature of tumors caused by disruption of the Drosophila epigenetic regulator, polyhomeotic (ph). In larval tissue ph mutant cells show a shift towards an embryonic-like signature. Using loss- and gain-of-function experiments we uncovered the embryonic transcription factor knirps (kni) as a new oncogene. The oncogenic potential of kni lies in its ability to activate JAK/STAT signaling and block differentiation. Conversely, tumor growth in ph mutant cells can be substantially reduced by overexpressing a differentiation factor. This demonstrates that epigenetically derailed tumor conditions can be reversed when targeting key players in the transcriptional network.

Effect of Sleep Duration, Diet, and Physical Activity on Obesity and Overweight Elementary School Students in Shanghai.

BACKGROUND: This was a cross-sectional survey to investigate the relationship of age, parent education, sleep duration, physical activity, and dietary habits with overweight or obesity in school-age children in Shanghai. METHODS: The survey gathered information from 13,001 children in grades 1 through 5 (age 6 to 10 years) among 26 elementary schools in 7 districts. Activity level was evaluated using the International Children's Leisure Activities Study Survey Questionnaire (CLASS-C). The definitions of normal, overweight, and obese were adjusted for each age. RESULTS: Logistic regression analysis indicated that age, being male, having ≤10 hours of sleep on non-school days, eating ≥1 vegetable/day, or drinking ≥1 sugar-sweetened drink/day increased the risk for a child being overweight or obese compared with having >10 hours of sleep or ≤3 vegetables or ≤3 sugar-sweetened drinks/month (p ≤ .008). Having >2 hours of outdoor activities on non-school days reduced the risk of being overweight or obese compared with ≤2 hours of outdoor activities on non-school days (p < .001). CONCLUSIONS: We found that age, sex, sleep, and some dietary habits impacted weight, and suggests that specific cultural and economic factors may impact risk of a child being overweight or obese.

The splicing co-factor Barricade/Tat-SF1 is required for cell cycle and lineage progression in Drosophila neural stem cells.

Stem cells need to balance self-renewal and differentiation for correct tissue development and homeostasis. Defects in this balance can lead to developmental defects or tumor formation. In recent years, mRNA splicing has emerged as an important mechanism regulating cell fate decisions. Here we address the role of the evolutionarily conserved splicing co-factor Barricade (Barc)/Tat-SF1/CUS2 in Drosophila neural stem cell (neuroblast) lineage formation. We show that Barc is required for the generation of neurons during Drosophila brain development by ensuring correct neural progenitor proliferation and differentiation. Barc associates with components of the U2 small nuclear ribonucleoprotein (snRNP) complex, and its depletion causes alternative splicing in the form of intron retention in a subset of genes. Using bioinformatics analysis and a cell culture-based splicing assay, we found that Barc-dependent introns share three major traits: they are short, GC rich and have weak 3' splice sites. Our results show that Barc, together with the U2 snRNP complex, plays an important role in regulating neural stem cell lineage progression during brain development and facilitates correct splicing of a subset of introns.

A regulatory transcriptional loop controls proliferation and differentiation in Drosophila neural stem cells.

Neurogenesis is initiated by a set of basic Helix-Loop-Helix (bHLH) transcription factors that specify neural progenitors and allow them to generate neurons in multiple rounds of asymmetric cell division. The Drosophila Daughterless (Da) protein and its mammalian counterparts (E12/E47) act as heterodimerization factors for proneural genes and are therefore critically required for neurogenesis. Here, we demonstrate that Da can also be an inhibitor of the neural progenitor fate whose absence leads to stem cell overproliferation and tumor formation. We explain this paradox by demonstrating that Da induces the differentiation factor Prospero (Pros) whose asymmetric segregation is essential for differentiation in one of the two daughter cells. Da co-operates with the bHLH transcription factor Asense, whereas the other proneural genes are dispensible. After mitosis, Pros terminates Asense expression in one of the two daughter cells. In da mutants, pros is not expressed, leading to the formation of lethal transplantable brain tumors. Our results define a transcriptional feedback loop that regulates the balance between self-renewal and differentiation in Drosophila optic lobe neuroblasts. They indicate that initiation of a neural differentiation program in stem cells is essential to prevent tumorigenesis.

[Multi-center study of premature thelarche and gynecomastia in Chinese infants and toddlers].

OBJECTIVE: The term "premature thelarche" refers to isolated breast development before 8 years of age in female, without any other signs of sexual maturation, while "gynecomastia" is the presence of breast tissue in males. This study aimed to investigate the prevalence of premature thelarche and gynecomastia in Chinese infants and toddlers, identify the potential risk factors, and explore the influence of early breast development on physical growth, mental development and psychomotor development. METHOD: A total of 1 510 full term and healthy children at the age of 0-48 months were sampled by stratified cluster random sampling method from 8 provinces from 2011-2012. Weight, height and breast development were assessed by senior primary pediatricians, while Bayley Scale of Infant Development-I (BSID-I) was used to measure the mental developmental index (MDI) and psychomotor developmental index (PDI) for children aged 2-30 months. Social-demographic Questionnaires were completed by the caregivers. RESULT: The combined prevalence of premature thelarche and gynecomastia was 1.6% (23/1 475), girls 2.2% (15/695), boys 1.0% (8/780), all within 2 years of age. The birth weight, feeding patterns in first 4 months, delivery mode, weaning time and social economic status were not significantly associated with the breast development. However, lower father's education level (OR = 3.632, 95%CI = 1.565-8.432) as well as smoking mother (OR = 18.960, 95%CI = 1.590-226.304) were significantly related to breast development even after adjusting for potential confounders. Lower weight (-0.479 ± 0.648 vs. 0.005 ± 0.987, P < 0.05) and height (-0.602 ± 1.042 vs. 0.008 ± 0.986, P < 0.05) Z score were found in breast development group, even after adjusting for age, gender and father' education level. Neither mental development (t = -0.082, P > 0.05) nor psychomotor development (t = 1.054, P > 0.05) was associated with breast development. CONCLUSION: We showed a similar prevalence of premature thelarche with the data reported in similar studies reported from other countries. Among the 0-48 months old infants and toddlers, Father's education level and smoking mother were both related to breast development. Breast development was significantly associated with physical growth, but had no correlation with the mental or psychomotor development.

Drosophila neural stem cells in brain development and tumor formation.

Neuroblasts, the neural stem cells in Drosophila, generate the complex neural structure of the central nervous system. Significant progress has been made in understanding the mechanisms regulating the self-renewal, proliferation, and differentiation in Drosophila neuroblast lineages. Deregulation of these mechanisms can lead to severe developmental defects and the formation of malignant brain tumors. Here, the authors review the molecular genetics of Drosophila neuroblasts and discuss some recent advances in stem cell and cancer biology using this model system.

SWI/SNF complex prevents lineage reversion and induces temporal patterning in neural stem cells.

Members of the SWI/SNF chromatin-remodeling complex are among the most frequently mutated genes in human cancer, but how they suppress tumorigenesis is currently unclear. Here, we use Drosophila neuroblasts to demonstrate that the SWI/SNF component Osa (ARID1) prevents tumorigenesis by ensuring correct lineage progression in stem cell lineages. We show that Osa induces a transcriptional program in the transit-amplifying population that initiates temporal patterning, limits self-renewal, and prevents dedifferentiation. We identify the Prdm protein Hamlet as a key component of this program. Hamlet is directly induced by Osa and regulates the progression of progenitors through distinct transcriptional states to limit the number of transit-amplifying divisions. Our data provide a mechanistic explanation for the widespread tumor suppressor activity of SWI/SNF. Because the Hamlet homologs Evi1 and Prdm16 are frequently mutated in cancer, this mechanism could well be conserved in human stem cell lineages. PAPERCLIP: