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Background Desmoplastic stroma, a feature of pancreatic ductal adenocarcinoma (PDAC), contains abundant activated pancreatic stellate cells (PSCs). How PSCs promote PDAC progression remains incompletely understood. Methods Effect of epithelium-specific E-twenty six factor 3 (ESE3)-positive PSCs on PDAC fibrosis and chemoresistance was examined by western blot, RT-PCR, immunofluorescence, flow cytometry assay, chromatin immunoprecipitation, luciferase assay, immunohistochemistry and subcutaneous pancreatic cancer mouse model. Results ESE3 expression increased in PSCs in PDAC tissues compared with those in normal PSCs. Clinical data showed that ESE3 upregulation in PSCs was positively correlated with tumour size, pTNM stage, CA19-9, carcinoembryonic antigen and serum CA242 level. ESE3 overexpression in PSCs was an independent negative prognostic factor for disease-free survival and overall survival amongst patients with PDAC. Mechanistically, the conditional medium from the loss and gain of ESE3-expressing PSCs influenced PDAC chemoresistance and tumour growth. ESE3 directly induced the transcription of α-SMA, collagen-I and IL-1β by binding to ESE3-binding sites on their promoters to activate PSCs. IL-1β upregulated ESE3 in PSCs through NF-κB activation, and ESE3 was required for PSC activation by tumour cell-derived IL-1β. Conclusion Inhibiting the IL-1β/ESE3 (PSCs)/IL-1β-positive feedback loop is a promising therapeutic strategy to reduce tumour fibrosis and increase chemotherapeutic efficacy in PDAC.

Background Chemoresistance is the main reason for the poor prognosis of pancreatic ductal adenocarcinoma (PDAC). Thus, there is an urgent need to screen out new targets and compounds to reverse chemotherapeutic resistance. Methods We established a bio-bank of human PDAC organoid models, covering a representative range of PDAC tumor subtypes. We screened a library of 1304 FDA-approved compounds to identify candidates efficiently overcoming chemotherapy resistance. The effects of the compounds were evaluated with a CellTiter-Glo-3D assay, organoid apoptosis assay and in vivo patient-derived xenograft (PDX), patient-derived organoid (PDO) and LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx1-Cre (KPC) genetically engineered mouse models. RNA-sequencing, genome editing, sphere formation assays, iron assays and luciferase assays were conducted to elucidate the mechanism. Results High-throughput drug screening of chemotherapy-resistant PDOs identified irbesartan, an angiotensin ‖ type 1 (AT1) receptor antagonist, which could synergistically enhance the ability of chemotherapy to kill PDAC cells. In vitro and in vivo validation using PDO, PDX and KPC mouse models showed that irbesartan efficiently sensitized PDAC tumors to chemotherapy. Mechanistically, we found that irbesartan decreased c-Jun expression by inhibiting the Hippo/YAP1 pathway and further overcame chemotherapy resistance in PDAC. We also explored c-Jun, a potential target of irbesartan, which can transcriptionally upregulate the expression of key genes involved in stemness maintenance (SOX9/SOX2/OCT4) and iron metabolism (FTH1/FTL/TFRC). More importantly, we observed that PDAC patients with high levels of c-Jun expression demonstrated poor responses to the current standard chemotherapy regimen (gemcitabine plus nab-paclitaxel). Moreover, patients with PDAC had significant survival benefits from treatment with irbesartan plus a standard chemotherapy regimen in two-center retrospective clinical cohorts and patients with high c-Jun expression exhibited a better response to combination chemotherapy. Conclusions Irbesartan could be used in combination with chemotherapy to improve the therapeutic efficacy in PDAC patients with high levels of c-Jun expression. Irbesartan effectively inhibited chemotherapy resistance by suppressing the Hippo/YAP1/c-Jun/stemness/iron metabolism axis. Based on our findings, we are designing an investigator-initiated phase II clinical trial on the efficacy and safety of irbesartan plus a standard gemcitabine/nab-paclitaxel regimen in the treatment of patients with advanced III/IV staged PDAC and are hopeful that we will observe patient benefits.

The ETS family transcription factor ESE3 is a crucial element in differentiation and development programs for many epithelial tissues. Here we report its role as a tumor suppressor in pancreatic cancer. We observed drastically lower ESE3 expression in pancreatic ductal adenocarcinomas (PDAC) compared with adjacent normal pancreatic tissue. Reduced expression of ESE3 in PDAC correlated closely with an increase in lymph node metastasis and vessel invasion and a decrease in relapse-free and overall survival in patients. In functional experiments, downregulating the expression of ESE3 promoted PDAC cell motility and invasiveness along with metastasis in an orthotopic mouse model. Mechanistic studies in PDAC cell lines, the orthotopic mouse model, and human PDAC specimens demonstrated that ESE3 inhibited PDAC metastasis by directly upregulating E-cadherin expression at the level of its transcription. Collectively, our results establish ESE3 as a negative regulator of PDAC progression and metastasis by enforcing E-cadherin upregulation. Cancer Res; 77(4); 874–85. ©2016 AACR.

Supplementary Data from Mitochondrial Calcium Uniporter Drives Metastasis and Confers a Targetable Cystine Dependency in Pancreatic Cancer

Pancreatic ductal adenocarcinoma (PDAC) is a highly metastatic disease with few effective treatments. Here we show that the mitochondrial calcium uniporter (MCU) promotes PDAC cell migration, invasion, metastasis, and metabolic stress resistance by activating the Keap1-Nrf2 antioxidant program. The cystine transporter SLC7A11 was identified as a druggable target downstream of the MCU-Nrf2 axis. Paradoxically, despite the increased ability to uptake cystine, MCU-overexpressing PDAC demonstrated characteristics typical of cystine-deprived cells and were hypersensitive to cystine deprivation-induced ferroptosis. Pharmacologic inhibitors of SLC7A11 effectively induced tumor regression and abrogated MCU-driven metastasis in PDAC. In patient-derived organoid models in vitro and patient-derived xenograft models in vivo, MCU-high PDAC demonstrated increased sensitivity to SLC7A11 inhibition compared with MCU-low tumors. These data suggest that MCU is able to promote resistance to metabolic stress and to drive PDAC metastasis in a cystine-dependent manner. MCU-mediated cystine addiction could be exploited as a therapeutic vulnerability to inhibit PDAC tumor growth and to prevent metastasis.Significance:Elevated mitochondrial calcium uptake in PDAC promotes metastasis but exposes cystine addiction and ferroptosis sensitivity that could be targeted to improve pancreatic cancer treatment.

Supplementary Table from Mitochondrial Calcium Uniporter Drives Metastasis and Confers a Targetable Cystine Dependency in Pancreatic Cancer

Figure S1. Effects of Methylation-reducing agent and ESE3 on pancreatic cancer growth and migration in vitro. Figure S2. The basal expression of ESE3 in four PDAC cell lines by Western blotting experiment. Figure S3. Regulation of EMT marker expression by ESE3. Figure S4. Impact of altered expression of ESE3 on pancreatic tumor growth in nude mice.

Purpose:Oxygen and glucose deprivation is a common feature of the solid tumor. Regulatory network underlying the adaptation of cancer cells to the harsh microenvironment remains unclear. We determined the mechanistic role of LIM and senescent cell antigen-like–containing domain protein 1 (LIMS1) in cancer cell survival under oxygen–glucose deprivation conditions.Experimental Design:The expression level of LIMS1 was determined by IHC staining and analyzing the mRNA expression profiles from The Cancer Genome Atlas of three human solid tumors. Roles of LIMS1 in cancer cell metabolism and growth were determined by molecular and cell biology methods. A jetPEI nanocarrier was used as the vehicle for anti-LIMS1 siRNAs in mouse models of cancer therapeutics.Results:LIMS1 expression was drastically elevated in pancreatic ductal adenocarcinoma (PDAC). High LIMS1 level was associated with advanced TNM stage and poor prognosis of patients with tumor. Increased LIMS1 expression was pivotal for tumor cells to survive in the oxygen–glucose deprivation conditions. Mechanistically, LIMS1 enhanced GLUT1 expression and membrane translocation, which facilitated tumor cell adaptation to the glucose deprivation stress. Furthermore, LIMS1 promoted HIF1A protein translation by activating AKT/mTOR signaling, while hypoxia-inducible factor 1 (HIF1) transactivated LIMS1 transcription, thus forming a positive feedback loop in PDAC cell adaptation to oxygen deprivation stress. Inhibition of LIMS1 with jetPEI nanocarrier–delivered anti-LIMS1 siRNAs significantly increased cell death and suppressed tumor growth.Conclusions:LIMS1 promotes pancreatic cancer cell survival under oxygen–glucose deprivation conditions by activating AKT/mTOR signaling and enhancing HIF1A protein translation. LIMS1 is crucial for tumor adaptation to oxygen–glucose deprivation conditions and is a promising therapeutic target for cancer treatment.

15 Supplementary Figures: Supplementary Figure S1. The expression and subcellular localization of LIMS1 in PDAC cell lines. Supplementary Figure S2. LIMS1 is overexpressed in esophageal carcinoma and negatively correlated with survival. Supplementary Figure S3. LIMS1 is overexpressed in lung adenocarcinoma and negatively correlated with survival. Supplementary Figure S4. LIMS1 had no effect on cell viability under normal culture condition. Supplementary Figure S5. LIMS1 facilitates glucose uptaking and surviving of esophageal carcinoma. Supplementary Figure S6. LIMS1 facilitates glucose uptaking and surviving of lung adenocarcinoma. Supplementary Figure S7. LIMS1 expression does not alter the half-life of HIF1A. Supplementary Figure S8. LIMS1 plays important roles in maintain the activation of AKT signaling. Supplementary Figure S9. Impact of altered expression of LIMS1 on tumorigenicity and therapeutic efficacy. Supplementary Figure S10. The effect of altered LIMS1 expression on extracellular acidification. Supplementary Figure S11. LIMS1 expression level positively correlates with glucose uptaking in PETCT scanning of patients with PDAC, esophageal carcinoma and lung adenocarcinoma. Supplementary Figure S12. The LIMS1-HIF1 positive feedback loop is pivotal to oxygen-glucose deprivation stress resistance. Supplementary Figure S13. Impact of altered expression on LIMS1 on cell survival under hypoxia. Supplementary Figure S14. Impact of altered expression of LIMS1 and HIF1 on tumorigenicity. Supplementary Figure S15. The effect of LIMS1 on IRES-dependent translation of HIF1A.

Figure S1. Effects of Methylation-reducing agent and ESE3 on pancreatic cancer growth and migration in vitro. Figure S2. The basal expression of ESE3 in four PDAC cell lines by Western blotting experiment. Figure S3. Regulation of EMT marker expression by ESE3. Figure S4. Impact of altered expression of ESE3 on pancreatic tumor growth in nude mice.

Supplementary Table 1. Antibodies and primers Supplementary Table 2. Clinicopathologic characteristics of TMA

7 Supplementary Tables: Supplementary Table S1. Association between LIMS1 expression and clinicopathological features of patients with PDAC. Supplementary Table S2. Association between LIMS1 mRNA expression and clinicopathological features of PDAC patients from TCGA. Supplementary Table S3. Cox's proportional hazards model analysis of prognostic factors in patients with pancreatic ductal adenocarcinoma (PDAC) Supplementary Table S4. Association between LIMS1 expression and clinicopathological features of patients with esophageal cancer. Supplementary Table S5. Association between LIMS1 expression and clinicopathological features of patients with lung adenocarcinoma. Supplementary Table S6. Main antibodies and reagents. Supplementary Table S7. Primers and oligonucleotides sequences.

Supplementary Table 1. Antibodies and primers.

Because of the early onset of local invasion and distant metastasis, pancreatic ductal adenocarcinoma (PDAC) is the most lethal human malignant tumor, with a 5-year survival rate of less than 5%. In this study, we investigated the role of fascin, a prometastasis actin-bundling protein, in PDAC progression, invasion, and the molecular mechanisms underlying fascin overexpression in PDAC. Our data showed that the expression levels of fascin were higher in cancer tissues than in normal tissues, and fascin overexpression correlated with the PDAC differentiation and prognosis. Fascin overexpression promoted PDAC cell migration and invasion by elevating matrix metalloproteinase-2 (MMP-2) expression. Fascin regulated MMP-2 expression through protein kinase C and extracellular signal—regulated kinase. Importantly, our data showed that hypoxia induced fascin overexpression in PDAC cells by promoting the binding of hypoxia-inducible factor-1 (HIF-1) to a hypoxia response element on the fascin promoter and transactivating fascin mRNA transcription. Intriguingly, HIF-1α expression levels in PDAC patient specimens significantly correlated with fascin expression. Moreover, immunohistochemistry staining of consecutive sections demonstrated colocalization between HIF-1α and fascin in PDAC specimens, suggesting that hypoxia and HIF-1α were responsible for fascin overexpression in PDAC. When ectopically expressed, fascin was able to rescue PDAC cell invasion after HIF-1α knockdown. Our results demonstrated that fascin is a direct target gene of HIF-1. Our data suggested that the hypoxic tumor microenvironment in PDAC might promote invasion and metastasis by inducing fascin overexpression, and fascin might be targeted to block PDAC progression. Cancer Res; 74(9); 2455–64. ©2014 AACR.

BxPC-3 and Panc-1 cells were transfected with pcDNA3.1 and pcDNA3.1-LASP1 plasmids (2 μg) for 48 h and assessed by RT-PCR and Western blot analyses.

(A) The morphology of Panc-1/pLV vector and Panc-1/pLV LASP-1 cells. (B) LASP-1 expression determined by Western blot analysis in Panc-1/pLV vector andPanc-1/pLV LASP-1 cells.

HIF-1α protein expression in CFPAC-1 cell transfected with negative control siRNA (siNC) and HIF-1α siRNA (si HIF-1α#1-3) (50 nM) for 48 hours determined by Western blot. β-actin was used as a loading control.

PDF file - 51K, Antibodies, siRNAs and primers.

Pancreatic ductal adenocarcinoma (PDAC) is a highly metastatic disease with few effective treatments. Here we show that the mitochondrial calcium uniporter (MCU) promotes PDAC cell migration, invasion, metastasis, and metabolic stress resistance by activating the Keap1-Nrf2 antioxidant program. The cystine transporter SLC7A11 was identified as a druggable target downstream of the MCU-Nrf2 axis. Paradoxically, despite the increased ability to uptake cystine, MCU-overexpressing PDAC demonstrated characteristics typical of cystine-deprived cells and were hypersensitive to cystine deprivation-induced ferroptosis. Pharmacologic inhibitors of SLC7A11 effectively induced tumor regression and abrogated MCU-driven metastasis in PDAC. In patient-derived organoid models in vitro and patient-derived xenograft models in vivo, MCU-high PDAC demonstrated increased sensitivity to SLC7A11 inhibition compared with MCU-low tumors. These data suggest that MCU is able to promote resistance to metabolic stress and to drive PDAC metastasis in a cystine-dependent manner. MCU-mediated cystine addiction could be exploited as a therapeutic vulnerability to inhibit PDAC tumor growth and to prevent metastasis. Significance: Elevated mitochondrial calcium uptake in PDAC promotes metastasis but exposes cystine addiction and ferroptosis sensitivity that could be targeted to improve pancreatic cancer treatment.

The store-operated calcium (Ca2+) entry (SOCE) is the Ca2+ entry mechanism used by cells to replenish depleted Ca2+ store. The dysregulation of SOCE has been reported in metastatic cancer. It is believed that SOCE promotes migration and invasion by remodeling the actin cytoskeleton and cell adhesion dynamics. There is recent evidence supporting that SOCE is critical for the spatial and the temporal coding of Ca2+ signals in the cell. In this review, we critically examined the spatiotemporal control of SOCE signaling and its implication in the specificity and robustness of signaling events downstream of SOCE, with a focus on the spatiotemporal SOCE signaling during cancer cell migration, invasion and metastasis. We further discuss the limitation of our current understanding of SOCE in cancer metastasis and potential approaches to overcome such limitation.

The covalently bound biomarkers in the macrostructure of kerogen or asphaltene can provide unique information compared with the routinely extractable biomarkers. Here, a shale sample (immature Posidonia Shale) and an oil sample (Norwegian Geochemical Standard NSO-1) were systematically investigated by heating them to different temperatures and by mixing them with different catalytic materials to optimize the microscale sealed vessel catalytic hydrogenation (MSSV-Hy) protocol. It can be demonstrated that (1) tetralin acts as an effective hydrogen donor to stabilize the generated free radicals in the pyrolysis system enabling the bound biomarkers to resemble more closely their free counterparts, (2) PtO2 is the most appropriate catalyst for promoting the overall release of bound biomarkers, and (3) 390 °C is the best end temperature (from 200 °C at 0.7 °C/min) for the MSSV-Hy approach, balancing between bound biomarker yields and the protection of the original biomarker steric configurations. MSSV-Hy is a quick, highly reproducible, less sample demanding, and cost-effective method to release bound biomarkers and has great potential for studying the carbon cycle in its broadest sense.

The crystal structure, electric properties, thermal stability and optical properties of Sm-doped 0.15Pb(Sc1/2Nb1/2)O3–0.50Pb(Mg1/3Nb2/3)O3–0.35PbTiO3 (xSm-doped PSN-PMN-PT) ternary multifunctional ceramics were investigated. It is found that the volume fraction of tetragonal phase increases with the increase in Sm3+ concentration. The ceramics with x = 0.005 and 0.01 show pure perovskite structure with coexistence of Mc and T phases. The optimized comprehensive properties are obtained in x = 0.01 ceramics: Tc, d33, kp are about 167 °C, 682 pC/N and 0.61, respectively. In the temperature range of 30–110 °C the variations in d33 and kp are only 5% and 9.8%, respectively. Meanwhile, excited by 410 nm, three distinct emission bands centered at 564 nm, 599 nm and 646 nm are detected and the highest intensity is detected in ceramics with x = 0.01. Our results indicate Sm-doped PSN-PMN-PT system is a promising multifunctional material for optical–electric devices applications.

Background Malnutrition and muscle wasting are common features frequently observed in pancreatic ductal adenocarcinoma (PDAC) patients with cancer cachexia. They are associated with reduced survival and quality of life. Nutrition therapy is an important part of multimodal cancer care in PDAC. However, due to the complexity of nutrition assessment, only 30–60% of patients with nutritional risks receive nutritional treatment at present. It is important to identify biomarkers that may be used to improve management of PDAC‐associated malnutrition. Serum insulin‐like growth factor binding protein 2 (IGFBP2) has emerged as a potential serum biomarker in a variety of tumours. However, its association with malnutrition and muscle wasting in PDAC is unclear. Methods We evaluated the tumour IGFBP2 expression and serum IGFBP2 level in 98 PDAC patients using immunohistochemistry and enzyme‐linked immunosorbent assay and analysed the correlation between them. Furthermore, we explored the relationship between IGFBP2 of both tumour and serum and nutritional status (Patient‐Generated Subjective Global Assessment and skeletal muscle index). Pan02 IGFBP2 stable transfection cell lines, Pan02 PLV‐IGFBP2 cells, and PLKO‐IGFBP2 cells were injected subcutaneously into the flank of C57BL/6 mouse. Serum IGFBP2 levels, food intake, and body weight of these mice were measured. The degree of muscle atrophy is characterized by haematoxylin and eosin, Oil Red O, and Masson's trichrome staining. The mRNA and protein expression of several essential muscle‐related signal proteins such as atrogin‐1 and muscle RING finger 1 was measured. Results Among 98 patients, we found that tumour IGFBP2 expression is related to plasma IGFBP2 levels (r s = 0.562, P

ObjectivePancreatic ductal adenocarcinoma (PDAC) is a highly lethal tumour with limited treatment options. Here, we identified syndecan binding protein (SDCBP), also known as syntenin1, as a novel targetable factor in promoting PDAC tumour progression. We also explored a therapeutic strategy for suppressing SDCBP expression.DesignWe used samples from patients with PDAC, human organoid models, LSL-KrasG12D/+mice, LSL-Trp53R172H/+ and Pdx1-Cre (KPC) mouse models, and PDX mouse models. Immunostaining, colony formation assay, ethynyl-2-deoxyuridine incorporation assay, real-time cell analysis, cell apoptosis assay, automated cell tracking, invadopodia detection and gelatin degradation assays, coimmunoprecipitation, and pull-down assays were performed in this study.ResultsThe median overall survival and recurrence-free survival rates in the high-SDCBP group were significantly shorter than those in the low-SDCBP group. In vitro and in vivo studies have demonstrated that SDCBP promotes PDAC proliferation and metastasis. Mechanically, SDCBP inhibits CK1δ/ε-mediated YAP-S384/S387 phosphorylation, which further suppresses β-TrCP-mediated YAP1 ubiquitination and proteasome degradation by directly interacting with YAP1. SDCBP interacts with the TAD domain of YAP1, mainly through its PDZ1 domain. Preclinical KPC mouse cohorts demonstrated that zinc pyrithione (ZnPT) suppresses PDAC tumour progression by suppressing SDCBP.ConclusionsSDCBP promotes the proliferation and metastasis of PDAC by preventing YAP1 from β-TrCP-mediated proteasomal degradation. Therefore, ZnPT could be a promising therapeutic strategy to inhibit PDAC progression by suppressing SDCBP.

Unlike the free biomarkers, which can be routinely recovered via solvent extraction, the bound biomarkers are covalently linked to the kerogen or asphaltene macrostructures and are more resistant to secondary alteration processes. Although similarities have been identified between these two types of biomarkers using different techniques, it remains unclear how the kerogen- and asphaltene-bound biomarkers are complicatedly corelated with their free counterparts, thus limiting the applications of bound biomarkers in geoscience research. In this study, the characteristics of free biomarkers recovered from shale samples, from 10 sedimentary basin around the world, and petroleum samples, from the North Sea, have been systematically compared with their kerogen- and asphaltene-bound biomarkers, which were released by the newly developed Microscale Sealed Vessel Catalytic Hydrogenation (MSSV-Hy) method. Although the kerogen-bound biomarkers are characterized by the absence of rearranged biomarkers (e.g., Ts and diasteranes) and elevated C29/C30 hopane ratios, many key source-related parameters show strong positive correlations between free and bound fractions, including the steranes/hopane, C27/C29 sterane, C30/C29 sterane, C29/C30 hopane (αβ + βα), C35/C34 homohopanes and C24 tetracyclic terpane/C26 tricyclic terpane ratios. These results suggest that bound biomarkers can reveal critical information about deposits and organic matter precursors of source rocks. Thermal maturity indicators, such as C29 sterane ββ/(αα + ββ), C29 sterane S/(S + R), C31 hopane S/R, and Tβ/Tm ratios, also display excellent correlations between the free and bound counterparts in the source rocks. Although the correlations are not 1:1, empirical formulae, which are based on the correlations between free and bound biomarkers, can be used to calibrate bound biomarkers into values that can be directly compared to routine free biomarkers. Importantly, asphaltene-bound biomarkers of North Sea samples exhibit similar characteristics to kerogen-bound biomarkers, and the conversion between free and bound biomarkers of the source rock and reservoir samples is the same. In summary, the bound biomarkers are found highly effective in determining the organic facies, lithology, and thermal maturity of the parent kerogen and/or asphaltene, and, thus, have a great potential to be widely used in geoscience research when information from free biomarkers is hampered by oil mixing, contamination or biodegradation.

Parvalbumin-expressing neurons are a type of inhibitory intermediate neuron that play an important role in terminating seizures. The aim of the present study was to use lentiviral construction and packaging technology to overexpress and silence the parvalbumin gene in pheochromocytoma (PC12) cells, and to evaluate how parvalbumin influences the metabolic pathway involving glutamate and γ-aminobutyric acid (GABA). In this work, Immunofluorescence staining was used to verify the differentiation of PC12 cells into neurons after adding nerve growth factor (NGF). Western blotting and real-time quantitative polymerase chain reaction (qRT-PCR) were used to confirm lentivirus-mediated knockdown or overexpression of parvalbumin. Expression of parvalbumin, the 65-kDa GAD isoform (GAD65), and the 67-kDa GAD isoform (GAD67) in neuronal cells was examined at the mRNA and protein levels using qRT-PCR, western blotting and immunofluorescence staining, while intracellular glutamate and GABA levels were determined by high performance liquid chromatography (HPLC). We demonstrate that the expression of parvalbumin is associated with GAD65 and GAD67. Interestingly, overexpression of parvalbumin up-regulated GAD65 and GAD67, increased GABA concentration, and decreased glutamate concentration. Silencing of parvalbumin led to the opposite effects. Altogether, parvalbumin affected the expression of GAD65 and GAD67, thereby influencing the metabolic pathway involving glutamate and GABA.

The very high temperature at the center of a hydrothermal system is detrimental for the deposition of organic-rich sediments. However, it's not known how the petroleum source rock develops at the boundary of a hydrothermal system where the seawater temperature was only slightly elevated. The petroleum generation potential, using pyrolysis-GC, GC-MS and FT-ICR MS techniques, of eight organic-rich Permian shale samples from Shangsi Section China were systematically investigated. Principal component analysis reveals that the quantity and quality of the organic matter in the shale is dominated by biomass productivity, the redox condition, and terrestrial organic matter input. Rare earth element distribution pattern reveals that the two samples with highest TOC contents show a slight Eu anomaly which indicates the sample was influenced by a hydrothermal system to some degree. A depositional model showing how the influences from hydrothermal event promote the development of petroleum source rock is summarized: (1) At the edge of a hydrothermal system, the seawater-temperature can be elevated by 5–10 °C which further promotes the development of phytoplankton and zooplankton. (2) An upwelling can be intrigued and chemical weathering will be accelerated which both introduce excessive nutrients and increase the biota productivity. (3) Flux with the terrestrial advanced plant can also be accelerated to the shelf area by the volcano-induced climate change. (4) The consumption of the extensive biomass induced by the hydrothermal activity would expand the oxygen minimum zone, and further enable more organic matter to be preserved in shale.

Desmoplastic stroma, a feature of pancreatic ductal adenocarcinoma (PDAC), contains abundant activated pancreatic stellate cells (PSCs). How PSCs promote PDAC progression remains incompletely understood.Effect of epithelium-specific E-twenty six factor 3 (ESE3)-positive PSCs on PDAC fibrosis and chemoresistance was examined by western blot, RT-PCR, immunofluorescence, flow cytometry assay, chromatin immunoprecipitation, luciferase assay, immunohistochemistry and subcutaneous pancreatic cancer mouse model.ESE3 expression increased in PSCs in PDAC tissues compared with those in normal PSCs. Clinical data showed that ESE3 upregulation in PSCs was positively correlated with tumour size, pTNM stage, CA19-9, carcinoembryonic antigen and serum CA242 level. ESE3 overexpression in PSCs was an independent negative prognostic factor for disease-free survival and overall survival amongst patients with PDAC. Mechanistically, the conditional medium from the loss and gain of ESE3-expressing PSCs influenced PDAC chemoresistance and tumour growth. ESE3 directly induced the transcription of α-SMA, collagen-I and IL-1β by binding to ESE3-binding sites on their promoters to activate PSCs. IL-1β upregulated ESE3 in PSCs through NF-κB activation, and ESE3 was required for PSC activation by tumour cell-derived IL-1β.Inhibiting the IL-1β/ESE3 (PSCs)/IL-1β-positive feedback loop is a promising therapeutic strategy to reduce tumour fibrosis and increase chemotherapeutic efficacy in PDAC.

Pancreatic cancer is a dismal malignancy with poor prognosis. In spite of progress in surgical technology, chemotherapy is still the cornerstone in the multi-disciplinary treatment. Albumin-bound paclitaxel is a first-line treatment for PDAC patients. Yet the response rate of the drug is far from satisfying. SOX8 is a member of the sex determining region Y-boxes family, which is potentially related to the chemoresistance of tumor. Patient with high expression of SOX8 were insensitive to albumin-bound paclitaxel. SOX8 reduced apoptosis and G2/M cell cycle arrest caused by albumin-bound paclitaxel. SOX8 transcriptionally regulated EZH2, which reduced expression of SPARC by promoting the methylation of SPARC, thereby reducing the transport of albumin-bound paclitaxel in pancreatic cancer cells. EZH2 inhibitor, UNC1999, can reverse the effect of SOX8 on chemo-resistance of albumin-bound paclitaxel. Collectively, our data revealed SOX8/EZH2/SPARC signaling induced primary chemo-resistance of albumin-bound paclitaxel in pancreatic ductal adenocarcinoma.

CD73, a cell surface-localized ecto-5'-nucleotidase, is the major enzymatic source of extracellular adenosine. Canonically, it plays multiple roles in cancer-related processes via its metabolite. As a druggable target, clinical trials targeting CD73 in various malignant diseases are currently ongoing. Here, we report the ecto-5'-nucleotidase-independent functions of CD73 in pancreatic ductal adenocarcinoma (PDAC). Our findings support that the elevated expression of CD73 in PDAC cells promotes gemcitabine (GEM) resistance by activating AKT. We discovered that a large amount of intracellular CD73 are localized in the endoplasmic reticulum membrane. Intracellular CD73 physically interacts with major vault protein to activate the SRC-AKT circuit. Troglitazone (TGZ) is a peroxisome proliferator-activated receptor gamma agonist that could inhibit the expression of CD73. The administration of TGZ markedly enhances sensitivity to GEM via downregulating CD73 in PDAC. Our findings support that CD73 could be targeted to overcome chemoresistance in PDAC.

Purpose: Oxygen and glucose deprivation is a common feature of the solid tumor. Regulatory network underlying the adaptation of cancer cells to the harsh microenvironment remains unclear. We determined the mechanistic role of LIM and senescent cell antigen-like–containing domain protein 1 (LIMS1) in cancer cell survival under oxygen–glucose deprivation conditions. Experimental Design: The expression level of LIMS1 was determined by IHC staining and analyzing the mRNA expression profiles from The Cancer Genome Atlas of three human solid tumors. Roles of LIMS1 in cancer cell metabolism and growth were determined by molecular and cell biology methods. A jetPEI nanocarrier was used as the vehicle for anti-LIMS1 siRNAs in mouse models of cancer therapeutics. Results: LIMS1 expression was drastically elevated in pancreatic ductal adenocarcinoma (PDAC). High LIMS1 level was associated with advanced TNM stage and poor prognosis of patients with tumor. Increased LIMS1 expression was pivotal for tumor cells to survive in the oxygen–glucose deprivation conditions. Mechanistically, LIMS1 enhanced GLUT1 expression and membrane translocation, which facilitated tumor cell adaptation to the glucose deprivation stress. Furthermore, LIMS1 promoted HIF1A protein translation by activating AKT/mTOR signaling, while hypoxia-inducible factor 1 (HIF1) transactivated LIMS1 transcription, thus forming a positive feedback loop in PDAC cell adaptation to oxygen deprivation stress. Inhibition of LIMS1 with jetPEI nanocarrier–delivered anti-LIMS1 siRNAs significantly increased cell death and suppressed tumor growth. Conclusions: LIMS1 promotes pancreatic cancer cell survival under oxygen–glucose deprivation conditions by activating AKT/mTOR signaling and enhancing HIF1A protein translation. LIMS1 is crucial for tumor adaptation to oxygen–glucose deprivation conditions and is a promising therapeutic target for cancer treatment.

EHF transcriptionally inhibits the expressions of TGFβ1 and GM-CSF to decrease T reg cell and MDSC accumulation, making it a promising biomarker to evaluate the immune microenvironment in PDAC. EHF overexpression may improve the efficacy of checkpoint immunotherapy in PDAC., Pancreatic ductal adenocarcinoma (PDAC) is a highly immune-suppressive tumor with a low response rate to single checkpoint blockade therapy. ETS homologous factor (EHF) is a tumor suppressor in PDAC. Here, we report a novel function of EHF in pancreatic cancer immune microenvironment editing and efficacy prediction for anti-PD1 therapy. Our findings support that the deficiency of tumoral EHF induced the accumulation of regulatory T (T reg) cells and myeloid-derived suppressor cells (MDSCs) and a decrease in the number of tumor-infiltrating CD8+ T cells. Mechanistically, EHF deficiency induced the conversion and expansion of T reg cells and MDSCs through inhibiting tumor TGFβ1 and GM-CSF secretion. EHF suppressed the transcription of TGFB1 and CSF2 by directly binding to their promoters. Mice bearing EHF overexpression tumors exhibited significantly better response to anti-PD1 therapy than those with control tumors. Our findings delineate the immunosuppressive mechanism of EHF deficiency in PDAC and highlight that EHF overexpression may improve PDAC checkpoint immunotherapy.

The organic matter- and uranium-rich Tremadocian Koporie Formation (“Dictyonema Shale” as traditional informal unit; 6–14 wt.% TOC, 74–274 ppm uranium) was studied with respect to the spatial and temporal distribution of the uranium contents in the shale since deposition. It was the aim to unravel organic-inorganic interactions as results of α-particle irradiation due to the decay of 238U and the consequences for the differential alteration of the organic matter and petroleum potential. A multi-method approach was applied to gain data on high resolution from electron microscopy, organic petrography, and organic and inorganic geochemistry about core samples from a borehole drilled 50 km southwest to St. Petersburg (Russia). The analytical results indicate that there were two sources of uranium. Bottom water anoxia promoted the reduction of soluble U(VI) from seawater (source 1) to U(IV) followed by adsorption in the sediment. This uranium portion is neither quantifiable on minerals nor on organic matter by EDX-SEM. Bottom water currents caused intercalations of coarser-grained intervals during deposition of the lower part of the Koporie Formation. These intervals include detrital uranium-rich apatite and zircon grains together with graptolite rhabdosomes which are also rich in uranium (source 2). Flushing of these permeable layers by meltwater during the Pleistocene (de)glaciation caused mobilization of U(VI) due to oxidation of formerly adsorbed quadrivalent uranium. The present uranium contents in the lower Koporie Formation are thus residues of formerly higher contents and indicate that stronger irradiation-induced destruction of the organic matter occurred during higher uranium contents before meltwater leaching during the Pleistocene. Today the organic matter of the lower part exhibits characteristics of a kerogen type III from the perspective of pyrolytic products with a rather Gas and Condensate potential whereas the upper part of the Koporie Formation still has the inherited signal of a kerogen type I/II with a Low-Wax Paraffinic Oil potential. Lithological and mineralogical heterogeneities in the lower part with locally strong irradiation are an alternative explanation for the stronger organic matter alteration.

Organic matter (OM) is known to be an important reductant in many sediment-hosted base metal deposits. However, the precise nature of interactions between OM and hydrothermal fluids are still debated, as well as how the interconnected reactions develop over geological timescales. We investigated the Kupferschiefer mineralization in two drill cores from the Spremberg-Graustein field in Eastern Germany focusing on the inventory of organonitrogen, -sulfur and -oxygen (NSO) compounds, which are most affected by organic-inorganic interactions with mineral surfaces, metals and aqueous fluids. Rather homogeneous deposition of type II marine organic matter, prevailing photic zone euxinia and an early oil window thermal maturity were documented using conventional hydrocarbon analysis. Furthermore, a slight level of hydrothermal alteration was detected in the lower part of drill core 121h/72, in proximity to the epigenetic hematitic alteration zone known as ‘Rote Fäule’: A minor loss of extractable organic matter and especially long-chain n-alkanes was accompanied by significant enrichment in oxygenated compounds and increased molecular oxygen content detected by FT–ICR–MS. A strong enrichment of deuterium was observed in the n-alkanes (up to -36‰) and isoprenoids (up to -20‰) indicate that hydrogen exchange with an evaporitic fluid occured in the Spremberg Kupferschiefer over geological timescales at maximum temperatures not exceeding 110 °C. We therefore conclude that FT–ICR–MS combined with the stable hydrogen isotope composition can provide valuable information about interaction time scales and the origin of mineralizing hydrothermal fluids.

To reveal the impact of biomass input (marine algae, terrestrial plants and lacustrine Botryococcus braunii) on the composition of high molecular weight organooxygen and organonitrogen compounds (Ox, Ny and NyOx), solvent extracts of immature–early mature rock and coal samples were systematically characterized by Fourier transform ion cyclotron resonance mass spectrometry using different ionization modes. In total, 16 samples from the marine Schöneck, Dynow and Posidonia formations, the lacustrine Wealden Formation and the terrestrial New Zealand coals were investigated. Coals as the in-situ deposits of terrestrial plants consist to a great degree of aromatic polyoxygenated Ox and N1Ox compounds, representing the degradation products of lignin like phenolic ketones and carboxylic acids as well as their condensation products with the degradation intermediates of labile organonitrogen compounds. Aliphatic Ox moieties derived from plant protective constituents principally waxes and cutan show a pronounced even or odd carbon number predominance of the C23–C33 species with C26, 28, 30 or C27, 29, 31 as the major homologs. In contrast, marine and lacustrine microbial communities contribute plentiful middle-chain C22, C24 or C23, C25 Ox compounds. Marine rock extracts are characterized by abundant organonitrogen compounds, especially N2 and N2Ox classes, interpreted as signatures of protein-rich marine algae that were successfully preserved via a degradation-recondensation pathway, whereas the low protein content of lacustrine Botryococcus braunii is reflected by a low amount of N1Ox and N1 classes. Highly aliphatic algaenan of Botryococcus braunii sterically protected its oxygen-bearing groups from degradation, leading to a great abundance of Ox compounds, furthermore, it characterizes the Botryococcus braunii source by substantial heteroatomic species containing more than 40 carbon atoms.

Fascin is a pro-metastasis actin bundling protein upregulated in essentially all the metastatic carcinoma. It is believed that fascin promotes cancer cell migration and invasion by facilitating membrane protrusions such as filopodia and invadopodia. Aerobic glycolysis is a key feature of cancer metabolism that provides critical intermediate metabolites for tumor growth and cell proliferation. Here we report that fascin increase glycolysis in lung cancer to promote tumor growth and metastasis. Fascin promotes glycolytic flux by increasing the expression and activities of phosphofructose kinase 1 and 2 (PFK1 and 2). The glycolytic function of fascin depends on activation of YAP1 through its canonical actin bundling activity. Fascin promotes the binding of YAP1 to a TEAD1/4 binding motif located 30 bp upstream of the PFKFB3 transcription start site to activate its transcription. Our interrogation of the TCGA database suggest that the fascin-YAP1-PFKFB3 circuit is likely conserved across different types of cancer. We further showed that the glycolytic function of fascin is essential for promotion of lung cancer growth and metastasis. Importantly, pharmacological inhibitors of fascin could be used to suppress the YAP1-PFKFB3 signaling and inhibit glycolysis in cancer cell lines, organoid cultures and xenograft metastasis models. Taken together, our data reveal an important glycolytic role of fascin in lung cancer metabolism, and suggest that pharmacological inhibitors of fascin could be used to reprogram cancer metabolism in lung cancer and potentially other cancer with fascin upregulation.

SummarySpaceflight has been documented to produce detrimental effects to physiology and genomic stability, partly a result of Galactic Cosmic Radiation (GCR). In recent years, extensive research into extremotolerant organisms has begun to reveal how they survive harsh conditions, such as ionizing radiation. One such organism is the tardigrade (Ramazzottius varieornatus) which can survive up to 5kGy of ionizing radiation and the vacuum of space. In addition to their extensive network of DNA damage response mechanisms, the tardigrade also possesses a unique damage suppressor protein (Dsup) that co-localizes with chromatin in both tardigrade and transduced human cells to protect against DNA damage from reactive oxygen species induced by ionizing radiation. While Dsup has been shown to confer human cells with increased radiotolerance; much of the mechanism of how it does this in the context of human cells remains unknown. Until now there is no knowledge yet of how introduction of Dsup into human cells can perturb molecular networks and if there are any systemic risks associated with foreign gene introduction. Here, we created a stable HEK293 cell line expressing Dsup, validated its radioprotective phenotype, and performed multi-omic analyses across different time points and doses of radiation to delineate molecular mechanism of the radioprotection and assess molecular network pertubations. Dsup expressing human cells showed an enrichment for pathways seen in cells overexpressing HMGN1, a chromosomal architectural protein that has a highly similar nucleosome binding motif. As HMGN1 binding to nucleosomes promotes a less transcriptionally repressed chromatin state, we further explored the hypothesis that Dsup could behave similarly via ATAC-seq analysis and discovered overall selective differential opening and closing of the chromatin landscape. Cut&Run analysis further revealed global increases in histone post translational modifications indicative of open chromatin and global decreases in repressive marks, with Dsup binding preferentially towards promoter regions marked by H3K27ac and H3K4me3. We further validated some of the enriched pathways via in-vitro assays and revealed novel phenotypes that Dsup confers to human cells such as reduction in apoptosis, increased cell proliferation, and increased cell adhesion properties. Our analysis provides evidence that the Dsup protein in the context of HEK293 cells may behave as a chromatin architectural protein and that in addition to its nucleosome shielding effect, may confer radio-resistance via chromatin modulation. These results provide future insight into mitigating some of the major challenges involved with long term spaceflight as well as understanding some of the molecular architectural underpinnings that lead to radioresistant cancer phenotypes back home.

Activating KRAS mutations are present in 25% of human cancer. Although oncogenic Ras was deemed “undruggable” in the past, recent efforts led to the development of pharmacological inhibitors targeting the KRASG12C mutant, which have shown promise in early clinical trials. The development of allele-specific K-RasG12C inhibitors marked a new chapter in targeting oncogenic KRAS mutant in cancer. However, drug resistance against these new drugs will likely limit their efficacy in the clinic. Genome-wide approaches have been used to interrogate the mechanisms of resistance to K-RasG12C inhibitors, which would facilitate the development of therapeutics overcoming drug resistance. This article reviews the latest progress in resistance to K-RasG12C-targeted therapies and aims to provide insight in future research targeting drug resistance in cancer.

Pancreatic stellate cells (PSCs) play a pivotal role in pancreatic fibrosis and pancreatic ductal adenocarcinoma (PDAC) progression. The mechanisms controlling PSC activation is not completely understood. Here we investigated the role of ESE3 (Epithelium-Specific ETS factor 3) in PSC activation. We discovered that in PDAC patients ESE3 expression was increased in PSC while decreased in tumor cells. ESE3 overexpression in PSC promoted PSC activation. Condition medium from ESE3-overexprssing PSC promotes PDAC cell migration, chemoresistance, tumor growth and fibrosis. ESE3 directly induced the transcription of α-SMA, Collagen 1 and IL-1β by binding to ESE3 binding sites on their promoters to activate PSC. On the other hand, IL-1β upregulates ESE3 in PSC through NFκB activation and ESE3 is required for PSC activation by tumor cell derived IL-1β. Clinical data showed ESE3 upregulation in PSC was positively correlated with tumor size, pTNM stage, CA19-9, CEA and CA242 level in serum. ESE3 overexpression in PSC was an independent negative prognostic factor for disease-free survival and overall survival among PDAC patients. Inhibition of the IL-1β/ ESE3 (PSC)/ IL-1β positive feedback loop represents a promising therapeutic strategy to reduce tumor fibrosis and increase chemotherapeutic efficacy in PDAC.

Self-construal priming was devised to mimic the effects of chronic cross-cultural differences. Primes designed to activate independent/interdependent self-construals have been found to affect numerous culturally relevant outcomes. However, researchers have rarely checked precisely what these primes activated, nor tested their cross-cultural equivalence. We compared two common priming tasks, Similarities vs. Differences with Family and Friends (SDFF) and Sumerian Warrior Story (SWS), across seven dimensions of independence/interdependence among 118 British and 178 Chinese participants. The two tasks activated different combinations of self-construal dimensions. SWS showed a similar pattern of effects across cultures, whereas SDFF more strongly affected Chinese participants. Neither manipulation closely mimicked the pattern of pre-existing cross-cultural differences between samples. We propose researchers should develop more precisely targeted self-construal primes.