Search

Plant-endophyte associations play important roles in grassland agricultural ecology. Many studies indicate that endophytic fungi can promote host growth and reduce the biotic and abiotic stresses of host plants. Epichloe endophytes of the family Clavicitaceae are fungal symbionts of Pooideae grasses. These endophytes usually do not express any obvious host symptoms. Many Epichloe endophytes can endow hosts with increased abiotic stress tolerance as a result of enhanced growth, tillering, reproduction and nutrient acquisition, particularly under conditions of drought, cold, salt and nutrient deficiencies and thus, have important roles in pastoral agricultural systems. Animal-safe grass-endophyte associations that confer bio-protective properties for increased pasture persistence and productivity have been developed and commercialized. Artificial inoculation with symbiotic Epichloe microbes is an important technique for the creation of novel germplasm. Selected Epichloe strains can add value to some grass-based forage systems by providing both biotic and abiotic stress resistance. In addition, they can improve and strengthen grass physiological functions and plant vigor. The Epichloe bromicola WBE1 endophyte of Hordeum brevisubulatum played an important role in maintaining the growth of host grass by promoting nutrient absorption and maintaining the ionic balance under salt stress. E. bromicola WBE1 can produce peramine, an alkaloid produced by Epichloe species, that protects host grasses from herbivorous insects. However, this strain was unable to synthesize the alkaloids that are toxic to livestock, such as ergine, ergonovine, ergovaline or lolitrem B. Barley ( Hordeum vulgare ) is one of the most important cereal crops. The importance of barley to human and animal nutrition, and indeed to the foundation and maintenance of human civilization, is well documented. Traditional barley breeding for improved environmental stress tolerance has resulted in good varieties. However, conventional breeding techniques neglect the microorganisms in plants. In this study, animal-safe endophytic E. bromicola WBE1 isolated from wild barley ( H. brevisubulatum ) was artificially inoculated into cultivated hulled barley ( H. vulgare cv. Yangsimai No. 1) and hull-less barley ( H. vulgare var. nudum cv. Chaiqing No. 1), creating novel barley germplasm. Plants that were inoculated with endophytes and those that were free of endophytes were evaluated for growth in the field. The plant height, tiller numbers, biomass and the grain weight per plant of endophyte-inoculated hulled barley plants (Yangsimai No. 1) were 5%, 20%, 46% and 22% higher than those of the control hulled barley plants, respectively, and they matured about 5 days earlier. The tiller numbers, biomass and grain weight per plant of endophyte-inoculated hull-less barley plants (Chaiqing No. 1) were 29%, 37% and 28% higher, respectively, than those of the control hull-less barley plants in the field. However, there was no significant difference between inoculated hull-less barley plants and control hull-less barley plants in the plant height and growth period. A novel germplasm of barley containing the endophyte was successfully created. E. bromicola improves plant characteristics and the growth of host. The results presented here provide evidence that the inoculation of barley with E. bromicola may be used to improve its germplasm. E. bromicola can be successfully inoculated into other phylogenetically close host species, and this technique has the potential to improve the growth of Hordeae cereal crops and serve as a broad application to produce agriculturally useful synthetic novel symbioses in germplasm.

Objective: To investigate the effects of microRNA-142-3p (miR-142-3p) on the biological characteristics of pancreatic cancer cells and its mechanism.Methods: The expression of miR-142-3p and nuclea...

Background Canavalia rosea (Sw.) DC. (bay bean) is an extremophile halophyte that is widely distributed in coastal areas of the tropics and subtropics. Seawater and drought tolerance in this species may be facilitated by aquaporins (AQPs), channel proteins that transport water and small molecules across cell membranes and thereby maintain cellular water homeostasis in the face of abiotic stress. In C. rosea, AQP diversity, protein features, and their biological functions are still largely unknown. Results We describe the action of AQPs in C. rosea using evolutionary analyses coupled with promoter and expression analyses. A total of 37 AQPs were identified in the C. rosea genome and classified into five subgroups: 11 plasma membrane intrinsic proteins, 10 tonoplast intrinsic proteins, 11 Nod26-like intrinsic proteins, 4 small and basic intrinsic proteins, and 1 X-intrinsic protein. Analysis of RNA-Seq data and targeted qPCR revealed organ-specific expression of aquaporin genes and the involvement of some AQP members in adaptation of C. rosea to extreme coral reef environments. We also analyzed C. rosea sequences for phylogeny reconstruction, protein modeling, cellular localizations, and promoter analysis. Furthermore, one of PIP1 gene, CrPIP1;5, was identified as functional using a yeast expression system and transgenic overexpression in Arabidopsis. Conclusions Our results indicate that AQPs play an important role in C. rosea responses to saline-alkaline soils and drought stress. These findings not only increase our understanding of the role AQPs play in mediating C. rosea adaptation to extreme environments, but also improve our knowledge of plant aquaporin evolution more generally.

Although the role of the Hippo signaling pathway in development and tumorigenesis has been extensively studied in multiple organs, its role in ovarian follicle development remains largely unknown. Here, we report that Yes-Associated Protein 1 (YAP1), the major effector of Hippo signaling, is spatiotemporally expressed in ovarian granulosa cells and plays a critical role in the regulation of follicle development. We found that the active form of YAP1 (nuclear YAP1) was predominantly expressed in proliferative granulosa cells, whereas the inactive form of YAP1 (cytoplasmic YAP1) was mainly detected in luteal cells (terminally differentiated granulosa cells). Pharmacological inhibition of YAP1 activity disrupted mouse ovarian follicle development in vitro and in vivo. Foxl2 promoter–driven knockout of Yap1 in ovarian granulosa cells resulted in increased apoptosis of granulosa cells, decreased number of corpora lutea, reduced ovarian size, and subfertility in transgenic mice. However, Cyp19a1 promoter–driven knockout of Yap1 in differentiated granulosa cells of preovulatory follicles and luteal cells of corpora lutea had no effect on ovarian morphology and fertility. Mechanistic studies demonstrated that YAP1 interacted with epidermal growth factor receptor and TGF-β signaling pathways to regulate granulosa cell proliferation, differentiation, and survival. Results from this study identify YAP1 as a critical regulator of granulosa cell proliferation and differentiation. Balanced expression and activation of YAP1 is essential for follicle development and successful reproduction. YAP1 is a promising target for treatment of subfertility associated with abnormal granulosa cell function.—Lv, X., He, C., Huang, C., Wang, H., Hua, G., Wang, Z., Zhou, J., Chen, X., Ma, B., Timm, B. K., Maclin, V., Dong, J., Rueda, B. R., Davis, J. S., Wang, C. Timely expression and activation of YAP1 in granulosa cells is essential for ovarian follicle development.

Canavalia rosea (Sw.) DC is a perennial twining herb distributed in the semi-arid and saline–alkali areas of coastal regions and has evolved halotolerance. In this study, we present the first comprehensive survey of the metallothionein (MT) gene family in C. rosea. MT proteins belong to a family of low-molecular-weight polypeptides with a high content of cysteine residues, which have an affinity to bind with heavy metal ions. MTs also play important roles in stress responses as reactive oxygen species (ROS) scavengers. A total of six CrMTs were identified in the C. rosea genome and classified into four subgroups by phylogenetic analysis. An analysis of the cis-acting elements revealed that a series of hormone-, stress-, and development-related cis-acting elements were present in the promoter regions of CrMTs. The expression of CrMTs also showed habitat- and environmental stress-regulated patterns in C. rosea. CrMT overexpression in yeast enhanced tolerance to heavy metals and ROS, as well as high osmotic and alkalinity stress, which is consistent with their predicted roles as metal-chelating proteins and ROS scavengers. Our results indicate that the CrMT genes might contribute to the detoxification of plants to metals and provide marked tolerance against abiotic stress. The expression patterns of CrMTs in C. rosea also indicate that CrMTs play important roles in this species’ response to extreme environments on tropical islands and reefs, probably by improving the thermotolerance of C. rosea plants.

Circular RNAs (circRNAs) are widely expressed in highly diverged eukaryotes. Although circRNAs have been known for many years, their function remains unclear. Interaction with RNA-binding protein (RBP) to influence post-transcriptional regulation is considered to be an important pathway for circRNA function, such as acting as an oncogenic RBP sponge to inhibit cancer. In this study, we design a deep learning framework, CRPBsites, to predict the binding sites of RBPs on circRNAs. In this model, the sequences of variable-length binding sites are transformed into embedding vectors by word2vec model. Bidirectional LSTM is used to encode the embedding vectors of binding sites, and then they are fed into another LSTM decoder for decoding and classification tasks. To train and test the model, we construct four datasets that contain sequences of variable-length binding sites on circRNAs, and each set corresponds to an RBP, which is overexpressed in bladder cancer tissues. Experimental results on four datasets and comparison with other existing models show that CRPBsites has superior performance. Afterwards, we found that there were highly similar binding motifs in the four binding site datasets. Finally, we applied well-trained CRPBsites to identify the binding sites of IGF2BP1 on circCDYL, and the results proved the effectiveness of this method. In conclusion, CRPBsites is an effective prediction model for circRNA-RBP interaction site identification. We hope that CRPBsites can provide valuable guidance for experimental studies on the influence of circRNA on post-transcriptional regulation.

Plant metal tolerance proteins (MTPs) play key roles in heavy metal absorption and homeostasis in plants. By using genome-wide and phylogenetic approaches, the origin and diversification of MTPs from Canavalia rosea (Sw.) DC. was explored. Canavalia rosea (bay bean) is an extremophile halophyte with strong adaptability to seawater and drought and thereby shows specific metal tolerance with a potential phytoremediation ability. However, MTP genes in leguminous plants remain poorly understood. In our study, a total of 12 MTP genes were identified in C. rosea. Multiple sequence alignments showed that all CrMTP proteins possessed the conserved transmembrane domains (TM1 to TM6) and could be classified into three subfamilies: Zn-CDFs (five members), Fe/Zn-CDFs (five members), and Mn-CDFs (two members). Promoter cis-acting element analyses revealed that a distinct number and composition of heavy metal regulated elements and other stress-responsive elements existed in different promoter regions of CrMTPs. Analysis of transcriptome data revealed organ-specific expression of CrMTP genes and the involvement of this family in heavy metal stress responses and adaptation of C. rosea to extreme coral reef environments. Furthermore, the metal-specific activity of several functionally unknown CrMTPs was investigated in yeast. These results will contribute to uncovering the potential functions and molecular mechanisms of heavy metal absorption, translocation, and accumulation in C. rosea plants.

Canavalia rosea (bay bean), distributing in coastal areas or islands in tropical and subtropical regions, is an extremophile halophyte with good adaptability to seawater and drought. Late embryogenesis abundant (LEA) proteins typically accumulate in response to various abiotic stresses, including dehydration, salinity, high temperature, and cold, or during the late stage of seed development. Abscisic acid-, stress-, and ripening-induced (ASR) genes are stress and developmentally regulated plant-specific genes. In this study, we reported the first comprehensive survey of the LEA and ASR gene superfamily in C. rosea. A total of 84 CrLEAs and three CrASRs were identified in C. rosea and classified into nine groups. All CrLEAs and CrASRs harbored the conserved motif for their family proteins. Our results revealed that the CrLEA genes were widely distributed in different chromosomes, and all of the CrLEA/CrASR genes showed wide expression features in different tissues in C. rosea plants. Additionally, we introduced 10 genes from different groups into yeast to assess the functions of the CrLEAs/CrASRs. These results contribute to our understanding of LEA/ASR genes from halophytes and provide robust candidate genes for functional investigations in plant species adapted to extreme environments.

Clinical and experimental evidence indicates that tumour‐associated macrophages support cancer progression. Moreover, macrophage‐derived extracellular vesicles (EVs) are involved in pathogenesis of multiple cancers, yet the functions of molecular determinants in which have not been fully understood. Herein, we aim to understand whether macrophage modulates pancreatic ductal adenocarcinoma (PDAC) progression in an EV‐dependent manner and the underlying mechanisms. microRNA (miR)‐365 was experimentally determined to be enriched in the EVs from M2 macrophages (M2‐EVs), which could be transferred into PDAC cells. Using a co‐culture system, M2‐EVs could enhance the proliferating, migrating and invading potentials of PDAC cells, while inhibition of miR‐365 in M2‐EVs could repress these malignant functions. B‐cell translocation gene 2 (BTG2) was identified to be a direct target of miR‐365, while the focal adhesion kinase (F/ATP)‐dependent tyrosine kinase (AKT) pathway was activated by miR‐365. We further demonstrated that overexpression of BTG2 could delay the progression of PDAC in vitro, whereas by impairing BTG2‐mediated anti‐tumour effect, M2‐EV‐miR‐365 promoted PDAC progression. For validation, a nude mouse model of tumorigenesis was established, in which we found that targeting M2‐EV‐miR‐365 contributed to suppression of tumour growth. Collectively, M2‐EVs carry miR‐365 to suppress BTG2 expression, which activated FAK/AKT pathway, thus promoting PDAC development.

Background: Canavalia rosea (Sw.) DC. (bay bean) is an extremophile halophyte that is widely distributed in coastal areas of the tropics and subtropics. Seawater and drought tolerance in this species may be facilitated by aquaporins (AQPs), channel proteins that transport water and small molecules across cell membranes and thereby maintain cellular water homeostasis in the face of abiotic stress. In C. rosea, AQP diversity, protein features, and their biological functions are still largely unknown.Results: We describe the action of AQPs in C. rosea using evolutionary analyses coupled with promoter and expression analyses. A total of 37 AQPs were identified in the C. rosea genome and classified into five subgroups: 11 plasma membrane intrinsic proteins, 10 tonoplast intrinsic proteins, 11 Nod26-like intrinsic proteins, 4 small and basic intrinsic proteins, and 1 X-intrinsic protein. Analysis of RNA-Seq data and targeted qPCR revealed organ-specific expression of aquaporin genes and the involvement of some AQP members in adaptation of C. rosea to extreme coral reef environments. We also analyzed C. rosea sequences for phylogeny reconstruction, protein modeling, cellular localizations, and promoter analysis. Furthermore, one of PIP1 gene, CrPIP1;5, was identified as functional using a yeast expression system and transgenic overexpression in Arabidopsis.Conclusions: Our results indicate that AQPs play an important role in C. rosea responses to saline-alkaline soils and drought stress. These findings not only increase our understanding of the role AQPs play in mediating C. rosea adaptation to extreme environments, but also improve our knowledge of plant aquaporin evolution more generally.

Circular RNAs (circRNAs) are extensively expressed in cells and tissues, and play crucial roles in human diseases and biological processes. Recent studies have reported that circRNAs could function as RNA binding protein (RBP) sponges, meanwhile RBPs can also be involved in back-splicing. The interaction with RBPs is also considered an important factor for investigating the function of circRNAs. Hence, it is necessary to understand the interaction mechanisms of circRNAs and RBPs, especially in human cancers. Here, we present a novel method based on deep learning to identify cancer-specific circRNA&ndash, RBP binding sites (CSCRSites), only using the nucleotide sequences as the input. In CSCRSites, an architecture with multiple convolution layers is utilized to detect the features of the raw circRNA sequence fragments, and further identify the binding sites through a fully connected layer with the softmax output. The experimental results show that CSCRSites outperform the conventional machine learning classifiers and some representative deep learning methods on the benchmark data. In addition, the features learnt by CSCRSites are converted to sequence motifs, some of which can match to human known RNA motifs involved in human diseases, especially cancer. Therefore, as a deep learning-based tool, CSCRSites could significantly contribute to the function analysis of cancer-associated circRNAs.

The immediate plasma metabolism and development of chemo-resistance (single agent) severely hampers the clinical effectiveness of Sorafenib (SRF) in liver cancer therapy. MicroRNA27a inhibition is a promising biological strategy for breast cancer therapy. In this study, we aimed to prepare SRF and anti-miRNA27a-loaded anti-GPC3 antibody targeted lipid nanoparticles to enhance the therapeutic efficacy against liver cancers. In this study, we have employed a unique cationic switchable lipid (CSL) as a mean to encapsulate miRNA as well as to confer pH-responsiveness to the nanocarrier system. The G-S27LN was nanosized and offered a pH-responsive release of SRF from the carrier system and we have demonstrated the specific affinity of G-S27LN towards the GPC3-overexpressed HepG2 cancer cells. Anti-microRNA27a significantly increased the protein expression of FOXO1 and PPAR-γ which are crucial components involved in proliferation and apoptosis of tumor cells. Combination of SRF and anti-miRNA27a (G-S27LN) resulted in significantly lower cell viability with a marked increase in the apoptosis cell proportion compared to that of free SRF indicating the synergistic anticancer effect. Animal studies in liver cancer xenograft model demonstrated significant suppression of tumor burden, reduced tumor cell and elevated TUNEL positive apoptosis with no toxicity concerns in animals treated with G-S27LN formulation. The CSL-based G-S27LN efficiently co-delivered anti-microRNA27a and SRF and therefore represents a promising therapy to treat liver cancer. This study also brings forth a platform strategy for the effective treatment of number of other advanced cancers.

Aquaporins are channel proteins that facilitate the transmembrane transport of water and other small neutral molecules, thereby playing vital roles in maintaining water and nutrition homeostasis in the life activities of all organisms. Canavalia rosea, a seashore and mangrove-accompanied halophyte with strong adaptability to adversity in tropical and subtropical regions, is a good model for studying the molecular mechanisms underlying extreme saline-alkaline and drought stress tolerance in leguminous plants. In this study, a PIP2 gene (CrPIP2;3) was cloned from C. rosea, and its expression patterns and physiological roles in yeast and Arabidopsis thaliana heterologous expression systems under high salt-alkali and high osmotic stress conditions were examined. The expression of CrPIP2;3 at the transcriptional level in C. rosea was affected by high salinity and alkali, high osmotic stress, and abscisic acid treatment. In yeast, the expression of CrPIP2;3 enhanced salt/osmotic and oxidative sensitivity under high salt/osmotic and H2O2 stress. The overexpression of CrPIP2;3 in A. thaliana could enhance the survival and recovery of transgenic plants under drought stress, and the seed germination and seedling growth of the CrPIP2;3 OX (over-expression) lines showed slightly stronger tolerance to high salt/alkali than the wild-type. The transgenic plants also showed a higher response level to high-salinity and dehydration than the wild-type, mostly based on the up-regulated expression of salt/dehydration marker genes in A. thaliana plants. The reactive oxygen species (ROS) staining results indicated that the transgenic lines did not possess stronger ROS scavenging ability and stress tolerance than the wild-type under multiple stresses. The results confirmed that CrPIP2;3 is involved in the response of C. rosea to salt and drought, and primarily acts by mediating water homeostasis rather than by acting as an ROS transporter, thereby influencing physiological processes under various abiotic stresses in plants.

Dysfunction of the homeostasis-maintaining systems in specific cell types or tissues renders the organism susceptible to a range of diseases, including cancers. One of the emerging mechanisms for maintaining tissue homeostasis is cellular senescence. Here, we report that the Hippo pathway plays a critical role in controlling the fate of ovarian cells. Hyperactivation of Yes-associated protein 1 (YAP1), the major effector of the Hippo pathway, induces senescence in cultured primary human ovarian surface epithelial cells (hOSEs). Large tumor suppressor 2 (LATS2), the primary upstream negative regulator of YAP1, is elevated in both YAP1-induced and natural replicative-triggered senescence. Deletion of LATS2 in hOSEs prevents these cells from natural replicative and YAP1-induced senescence. Most importantly, loss of LATS2 switches ovarian cells from YAP-induced senescence to malignant transformation. Our results demonstrate that LATS2 and YAP1, two major components of the Hippo/YAP signaling pathway, form a negative feedback loop to control YAP1 activity and prevent ovarian cells from malignant transformation. Human cancer genomic data extracted from TCGA datasets further confirm the clinical relevance of our finding.

Seasonal precipitation changes are increasingly severe in subtropical areas. However, the responses of soil nitrogen (N) cycle and its associated functional microorganisms to such precipitation changes remain unclear. In this study, two projected precipitation patterns were manipulated: intensifying the dry-season drought (DD) and extending the dry-season duration (ED) but increasing the wet-season storms following the DD and ED treatment period. The effects of these two contrasting precipitation patterns on soil net N transformation rates and functional gene abundances were quantitatively assessed through a resistance index. Results showed that the resistance index of functional microbial abundance (-0.03 ± 0.08) was much lower than that of the net N transformation rate (0.55 ± 0.02) throughout the experiment, indicating that microbial abundance was more responsive to precipitation changes compared with the N transformation rate. Spring drought under the ED treatment significantly increased the abundances of both nitrifying (amoA) and denitrifying genes (nirK, nirS, and nosZ), while changes in these gene abundances overlapped largely with control treatment during droughts in the dry season. Interestingly, the resistance index of the ammonia-oxidizing archaea (AOA) amoA abundance was significantly higher than that of the denitrifying gene abundances, suggesting that AOA were more resistant to the precipitation changes. This was attributed to the stronger environmental adaptability and higher resource utilization efficiency of the AOA community, as indicated by the lack of correlations between AOA gene abundance and environmental factors [i.e., soil water content, ammonium (NH4+) and dissolved organic carbon concentrations] during the experiment.

Transient cerebral ischemia dramatically activates small ubiquitin-like modifier (SUMO2/3) conjugation. In cells exposed to 6 h of transient oxygen/glucose deprivation (OGD), a model of ischemia, SUMOylation increases profoundly between 0 and 30 min following re-oxygenation. To elucidate the effect of transient OGD on SUMO conjugation of target proteins, we exposed neuroblastoma B35 cells expressing HA-SUMO3 to transient OGD and used stable isotope labeling with amino acids in cell culture (SILAC) to quantify OGD-induced changes in levels of specific SUMOylated proteins. Lysates from control and OGD-treated cells were mixed equally, and HA-tagged proteins were immunoprecipitated and analyzed by 1D-SDS-PAGE-LC-MS/MS. We identified 188 putative SUMO3-conjugated proteins, including numerous transcription factors and coregulators, and PIAS2 and PIAS4 SUMO ligases, of which 22 were increased or decreased more than ±2-fold. In addition to SUMO3, the levels of protein-conjugated SUMO1 and SUMO2, as well as ubiquitin, were all increased. Importantly, protein ubiquitination induced by OGD was completely blocked by gene silencing of SUMO2/3. Collectively, these results suggest several mechanisms for OGD-modulated SUMOylation, point to a number of signaling pathways that may be targets of SUMO-based signaling and recovery from ischemic stress, and demonstrate a tightly controlled crosstalk between the SUMO and ubiquitin conjugation pathways.

The Hippo pathway plays critical roles in both organ development and tumorigenesis. Our previous studies demonstrated that Yes-Associated Protein 1 (YAP), a transcription coactivator and the major effector of the Hippo pathway, plays a critical role in ovarian cancer development. The present study investigates the role of the Hippo/YAP signaling pathway in the tumorigenesis of ovarian granulosa cells. Fluorescent and chromogenic immunohistochemistry were used to examine YAP expression in human and mouse ovaries. Western blot and qRT-PCR were used to determine protein and mRNA expression. Retrovirus-based YAP expressing constructs and YAP siRNAs were utilized to manipulate YAP levels in primary or immortalized human granulosa cells. A new three-dimensional hanging-drop culture system was used to mimic the growth of granulosa cells in vivo. The soft agar assay and xenograft mouse models were employed to investigate the role of YAP in transformation and tumorigenesis of granulosa cells. Immunohistochemical analyses showed that the active form of YAP (nuclear YAP) was expressed in proliferative granulosa cells in human and mouse ovaries, whereas the inactive form of YAP (cytoplasmic YAP) was detected predominantly in terminally differentiated luteal cells, indicating that YAP activity may be involved in the regulation of granulosa cell proliferation and differentiation. Overexpression of wild-type or constitutively active YAP compromised the expression of genes required for steroidogenesis and induced de-differentiation of differentiated granulosa cells. Ectopic expression of the constitutively active form of YAP in less-differentiated human granulosa cells derived from growing follicles induced tumors in a xenograft mouse model. Intriguingly, we found that these tumors resembled human high-grade serous ovarian cancer (HGSOC) morphologically, histologically, and genetically. The tumors were accompanied by accumulation of large amount of ascites in the peritoneal cavity. Highly proliferative and aggressive tumor cells invaded into omentum, visceral fat tissue, diaphragm, pancreas, liver, and gastrointestinal system. Deletion of the BRCA1 gene in these tumor cells promoted tumor progression and significantly reduced survival rate. Interestingly, these tumors have high expression of N-cadherin but very low expression of CA125 and E-cadherin, which are molecular characteristics of a recently identified mesenchymal subtype of HGSOC. Studies from The Cancer Genome Atlas and the Australian Ovarian Cancer Study Group indicate that the mesenchymal subtype of HGSOC has the poorest prognosis among all subtypes of ovarian cancers. In conclusion, results from this study indicate that YAP plays a critical role in regulating granulosa cell proliferation and differentiation. Overactivation of YAP can result in de-differentiation and reprogramming of ovarian granulosa cells, leading to development of mesenchymal subtype of HGSOC. Our results identify early-stage granulosa cells as a cell of origin of newly identified mesenchymal type of HGSOC. Citation Format: Xiangmin Lv, Chunbo He, Cong Huang, Guohua Hua, Bowen Ma, Xingcheng Chen, Jin Zhou, Zhengfeng Wang, Jixin Dong, Bo R. Rueda, John S. Davis, Cheng Wang. YAP induces development of mesenchymal subtype of high-grade serous ovarian cancer from granulosa cells. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr B50.

The herb Astragalus membranaceus is used in traditional Chinese medicine to boost immunity. This study investigated the effects of Astragalus polysaccharides (APS) and astragalosides (AS) on the phagocytosis of Mycobacterium tuberculosis by macrophages. Peritoneal macrophages were obtained by peritoneal lavage from mice stimulated by starch gravy culture medium and cultured with M. tuberculosis and varying concentrations of APS and AS. Phagocytotic activity was measured using a real-time polymerase chain reaction assay to detect M. tuberculosis DNA. Levels of interleukin-1β, interleukin-6 and tumour necrosis factor-α secreted by activated macrophages in the culture supernatant were determined using an enzyme-linked immunosorbent assay. Macrophage phagocytotic activity and secreted cytokine levels were significantly increased after treatment with APS and AS. This study provides evidence that APS and AS have strong promoting effects on the phagocytosis of M. tuberculosis by macrophages and the secretion of interleukin-1β, interleukin-6 and tumour necrosis factor-α by activated macrophages.

INTRODUCTION: Accumulating evidence indicates that ovarian high-grade serous carcinoma (HGSC) originates from Fallopian tube secretory epithelial cells (FTSECs). Our recent studies indicated that the Hippo/YAP signaling pathway contributes to transformation of ovarian surface epithelial cells. However, whether this pathway is also involved in the initiation and progression of FTSEC-derived ovarian HGSC has not been examined. AIMS: The present study aims to determine whether the Hippo/YAP signaling pathway plays a role in initiation and progression of Fallopian tube-derived HGSC and the potential mechanism(s) by which YAP regulates FTSEC tumorigenesis. RESULTS: Immunohistochemical analysis showed that YAP expression was very low in normal human fallopian tube and ovarian tissues. However, YAP immunosignals significantly increased in ovarian HGSCs, cancerous and inflammatory fallopian tube tissues. Data extracted from multidimensional human cancer genomics datasets revealed that the YAP and TEADs (the major target transcription factors of YAP) genes are frequently amplified and/or up-regulated, while the upstream tumor suppressor genes such as NF1 and LATS1, are frequently deleted and/or mutated in ovarian HGSC tissues. Overexpression of YAP in immortalized non-tumorigenic FTSECs promoted cell proliferation in both 2D and 3D culture systems, induced colony formation in soft agar, and drove tumorigenesis in athymic nude mice. Pathohistological and biochemical analyses clearly indicated that tumors derived from the FTSECs resembled ovarian HGSC. In addition, our mechanistic studies showed that activation of YAP in human FTSECs upregulated the expression of fibroblast growth factors (FGF1, FGF2) and their receptors (FGFR1, FGFR2, FGFR3 and FGFR4). Intriguingly, we found that the activated FGF signaling in turn suppressed the Hippo pathway and activated YAP protein. Consistent with this result, treatment of FTSECs with YAP antagonist (Verteporfin) and/or FGFR inhibitor (BGI398) not only blocked YAP-induced FTSECs transformation, but also suppressed tumor cell growth and induced tumor cell death in a human HGSC xenograft model. CONCLUSION: Our results demonstrate that the disruption of the Hippo/YAP signaling pathway contributes to the carcinogenesis of the fallopian tube secretory epithelial cells. The Hippo/YAP and FGF signaling pathways formed an autocrine/paracrine positive feedback loop to drive the progression of the FTSECs-derived HGSC. Evidence in this study strongly suggests that combined targeting of YAP and FGFRs represents a novel therapeutic strategy for ovarian HGSCs. Citation Format: Chunbo He, Guohua Hua, Xiangmin Lv, Zhengfeng Wang, Ronny I. Drapkin, John S. Davis, Cheng Wang. YAP, VIA INTERACTIONS WITH THE FGF SIGNALING PATHWAY, REGULATES TUMORIGENESIS OF FALLOPIAN TUBE SECRETORY EPITHELIAL CELLS [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr MIP-058.

The tumor suppressor p53 is a master regulator of apoptosis and also plays a key role in cell cycle progress and cell differentiation. It mainly acts as a transcription factor. In addition, it can also directly interact with apoptosis regulators in mitochondria to control apoptosis. Recent studies from our laboratory and others have shown that p53 plays an active role in regulating lens differentiation. It does so by modulating developmental apoptosis and also controlling expression of lens differentiation-specific genes. In this chapter, we summary the current progresses in this field.