Evidence Of Alterations In Expression Research Articles

Changes in the Coexpression of Innate Immunity Genes During Persistent Islet Autoimmunity Are Associated With Progression of Islet Autoimmunity: Diabetes Autoimmunity Study in the Young (DAISY).

Longitudinal changes in gene expression during islet autoimmunity (IA) may provide insight into biological processes that explain progression to type 1 diabetes (T1D). We identified individuals from Diabetes Autoimmunity Study in the Young (DAISY) who developed IA, autoantibodies present on two or more visits. Illumina's NovaSeq 6000 was used to quantify gene expression in whole blood. With linear mixed models we tested for changes in expression after IA that differed across individuals who progressed to T1D (progressors) (n = 25), reverted to an autoantibody-negative stage (reverters) (n = 47), or maintained IA positivity but did not develop T1D (maintainers) (n = 66). Weighted gene coexpression network analysis was used to identify coexpression modules. Gene Ontology pathway analysis of the top 150 differentially expressed genes (nominal P < 0.01) identified significantly enriched pathways including leukocyte activation involved in immune response, innate immune response, and regulation of immune response. We identified a module of 14 coexpressed genes with roles in the innate immunity. The hub gene, LTF, is known to have immunomodulatory properties. Another gene within the module, CAMP, is potentially relevant based on its role in promoting β-cell survival in a murine model. Overall, results provide evidence of alterations in expression of innate immune genes prior to onset of T1D.

Abstract 5175: Birinapant enhances death agonist antibody against TRAILR2 anti-tumor activity in HPV-positive head and neck squamous cell carcinomas

Abstract Human papilloma virus positive (HPV+) head and neck squamous cell carcinomas (HNSCC) exhibit a better prognosis and response to therapies than HPV(-) cancers. Analysis of HNSCC TCGA dataset provides evidence for distinct alterations in expression of components of the NF-κB and cell death pathways in HPV(+/-) HNSCC. Previously, we have found that birinapant, a novel SMAC mimetic that inhibits inhibitor of apoptosis proteins (IAPs), sensitizes a subset of HPV(-) HNSCC cell lines to death agonists like TNF-α and TRAIL. In our current study, we have observed that birinapant also sensitizes several HPV(+) cell lines to TNF-α and TRAIL in vitro. The IC50 for birinapant was under 50nM with TRAIL and TNF-α for the HPV (+) UPCI-SCC-90 and UM-SCC-47 cell lines. As TRAIL is known as a selective cancer cell death inducer, we explored its potential for enhancing death signaling in HPV(+) HNSCC cells via the effects of an agonistic polyclonal TRAILR2 antibody. Flow cytometry analysis confirmed the presence of TRAILR2 expression on the cell surface of the two HPV(+) cell lines. Treatment of cells with the antibody or TRAIL alone showed little or no inhibitory effect on growth of either cell lines. However, treatment with birinapant plus antibody, and especially the triple combination of birinapant, TRAIL, and antibody showed additive or synergistic effects to inhibit cell proliferation in a dose dependent manner. The Fixed Apoptotic Necrotic test using anti-phosphatidylserine and Zombie fixable dye showed that late apoptosis was the predominant mode of cell death for all combination groups for both cell lines at 48 hours post treatment. The proportion of cells undergoing apoptosis/necrosis was highest in the triple combination group in both cell lines, followed by birinapant and TRAIL combination for UM-SCC-47, or birinapant and antibody combination for UPCI-SCC-90, which was consistent with the relative percentage of sub-G0 DNA determined by flow cytometry analysis using propidium iodide staining. Enhanced apoptosis/necrosis with the triple combination was observed earlier than with single or dual treatment (12 versus 24 hours post treatment). Specific cell death mechanism was analyzed using pan-caspase (ZVAD), caspase-8 (ZIETD), and RIPK1 (necrostatin) inhibitors. In both UPCI-SCC-90 and UM-SCC-47 cells, caspase inhibition (ZVAD and ZIETD) completely reversed the effects of the double or triple combination treatments, whereas necrostatin did not. This suggests that the TRAILR2 agonist antibody mediated cell death in combination with birinapant is predominantly caspase-8 dependent. These results indicate that the TNF-α, TRAIL, and TRAILR2 agonist antibody sensitized birinapant anti-tumor activity, and triple combination exhibited synergistic effects. Supported by NIDCD projects ZIA-DC-000016, -73, -74, and the Medical Research Scholars Program. Citation Format: Yi An, Jun W. Jeon, Lillian Sun, Adeeb Derakhshan, Jianhong Chen, Hui Cheng, Xinping Yang, Christopher Silvin, Carter Van Waes, Zhong Chen. Birinapant enhances death agonist antibody against TRAILR2 anti-tumor activity in HPV-positive head and neck squamous cell carcinomas [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5175.

Abstract 3974: Combination of birinapant and TRAILR2 agonist antibody enhances cell death in HPV-positive head and neck squamous cell carcinomas

Abstract Head and neck squamous cell carcinomas (HNSCC) induced by human papilloma virus (HPV) have increased recently in the US. Analysis of HNSCC TCGA datasets provide evidence for distinct alterations in expression of NF-κB components and death pathways in HPV(+) HNSCC. Previously, we have found that Birinapant, a novel SMAC mimetic that inhibits inhibitor of apoptosis proteins (IAPs), exhibits single agent activity and sensitizes a subset of HPV(-) HNSCC cell lines to death agonists like TNF-α and TRAIL. Birinapant exhibited anti-tumor effects in vivo for HNSCC with FADD amplification. However, Birinapant alone exhibited minimal activity against HPV(+) tumor in vivo. To explore how to enhance the sensitivity of HPV(+) cells to Birinapant, we treated a panel of cell lines with Birinapant alone, and in combination with TRAIL or TNF-α in vitro. Birinapant sensitized HPV(+) cell lines UPCI-SCC-90 and UM-SCC-47 to TRAIL or TNF-α. The IC50 for Birinapant was under 50nM when combined with TRAIL or TNF-α for UPCI-SCC-90 and UM-SCC-47 cell lines. To explore the therapeutic potential of enhancing TRAILR mediated death signaling in HPV(+) HNSCC cells, we investigated the effects of an agonistic polyclonal TRAILR2 antibody. Anti-TRAILR2, TRAIL, or TNF-α alone showed little or no inhibitory effect on proliferation of UPCI-SCC-90 and UM-SCC-47 cells in vitro. However, a combination of Birinapant and TRAILR2 antibody, and triple combination of Birinapant, TRAIL, and TRAILR2 antibody showed additive or synergistic effects to inhibit cell density in a dose dependent manner. Combination of Birinapant with TRAIL, TRAILR2 antibody, or triple combination led to cell death and subG0 DNA fragmentation, as demonstrated by propidium iodide staining and flow cytometry. To further analyze the mechanism of cell death, we treated cells with the inhibitor of pan-caspase (ZVAD), caspase-8 (ZIETD), or RIPK1 (necrostatin) inhibitors. In both UPCI-SCC 90 and UM-SCC 47 cells, caspase inhibition (ZVAD and ZIETD) completely abolished the effects of Birinapant treatment in combination with TRAIL, TRAILR2 antibody, and triple combination, whereas RIPK1 inhibition (necrostatin) did not reverse the effects significantly. Our data suggest that Birinapant and TRAIL receptor mediated cell death is predominantly caspase 8 dependent. Taken together, our study indicates that TRAIL or TRAILR2 agonist antibody enhances Birinipant anti-tumor activity, and triple combination exhibits the strongest synergistic effects. The triple combination of targeting death pathway by TRAIL and TRAILR2 antibody with Birinapant could be a potent strategy to treat aggressive subset of HPV(+) HNSCC, and warrants the testing in future clinical trials. (Supported by NIDCD intramural projects ZIA-DC-000016, 74). Citation Format: Yi An, Lillian Sun, Adeeb Derakhshan, Sophie Carlson, Zhong Chen, Carter Van Waes. Combination of birinapant and TRAILR2 agonist antibody enhances cell death in HPV-positive head and neck squamous cell carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3974.

Abstract 1061: Modulation of death pathways by TRAILR2 agonist antibody and NF-κB pathway by NIK inhibitor in HPV-positive head and neck squamous cell carcinomas

Abstract Head and neck squamous cell carcinomas (HNSCC) induced by human papilloma virus (HPV) have increased recently in the US, and exhibit a different prognosis and response to therapies from HPV(-) cancers. Analysis of HNSCC TCGA datasets provide evidence for distinct alterations in expression of components of the NF-κB and death pathways in HNSCC with different HPV status. Previously, we have found that birinipant, a novel SMAC mimetic that inhibits inhibitor of apoptosis proteins (IAPs), sensitizes a subset of HPV(-) HNSCC cell lines to death agonists like TNF-α and TRAIL. In this study, we have observed that birinipant also sensitizes most HPV(+) cell lines to TRAIL and TNF-α in vitro. The IC50 of birinipant was under 50nM for HPV(+) UPCI-SCC-90 and UM-SCC-47 cell lines, when combined with TNF-α or TRAIL. To explore the therapeutic potential of enhancing TRAILR mediated death signaling in HPV(+) HNSCC cells, we investigated the effects of an agonistic polyclonal TRAILR2 antibody. Treatment of cells with TRAILR2 antibody alone shows little or no inhibitory effect on UPCI-SCC-90 and UM-SCC-47 cells in vitro. However, a combination of birinipant and TRAILR2 antibody, or triple combination of birinipant, TRAIL, and TRAILR2 antibody shows additive or synergistic effects to inhibit cell proliferation and induce cell death in a dose dependent manner. In addition, our preliminary data suggested that the non-canonical NF-κB pathway is predominately activated in HPV(+) HNSCC cells, and that NF-κB inducing kinase (NIK) is a key component of this pathway. When we tested the NIK inhibitor 4H-isoquinoline-1,3-dione in UPCI-SCC-90 cells, we found it reduced cell proliferation in a dose-dependent manner (IC50 =1.5 μM). Taken together, these results indicate that TNF-α, TRAIL, and TRAILR2 agonist antibody sensitized birinipant anti-tumor activity, and triple combination exhibited synergistic effects in HPV(+) HNSCC cell lines. NIK inhibitor alone inhibits cell proliferation of HPV(+) cells, supporting the hypothesis of aberrant activation of alternative pathway in HPV(+) HNSCC cells. (Supported by NIDCD intramural projects ZIA-DC-000016, 73, 74). Citation Format: Yi An, Lillian Sun, Adeeb Derakhshan, Sophie Carlson, Rita Das, Zhong Chen, Carter Van Waes. Modulation of death pathways by TRAILR2 agonist antibody and NF-κB pathway by NIK inhibitor in HPV-positive head and neck squamous cell carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1061. doi:10.1158/1538-7445.AM2017-1061

Neurotrophin receptors expression in mesial temporal lobe epilepsy with and without psychiatric comorbidities and their relation with seizure type and surgical outcome.

Epilepsy and psychiatric comorbidities are frequently associated, but their common biological substrate is unknown. We have previously reported altered structural elements and neurotrophins (NTs) expression in mesial temporal lobe epilepsy (MTLE) patients with psychiatric comorbidities. NTs receptors can regulate neurotransmission and promote neuroplasticity, being important candidates in the regulation and manifestation of psychopatological states and seizure-related events. MTLE hippocampi of subjects without psychiatric history, MTLE + major depression, MTLE + interictal psychosis derived from epilepsy surgery, and control necropsies were investigated for p75NTR, TrkB, TrkA, and TrkC immunohistochemistry. Increased expression of p75NTR, decreased TrkA, unaltered TrkC, and complex alterations involving TrkB expression were seen in MTLE groups. Increased TrkB expression in patients without complete seizure remission and in those with secondarily generalized seizures was seen. Decreased p75NTR expression associated with interictal psychosis, and increased TrkB in those with psychosis or major depression was also reported, although their p75NTR/TrkB ratios were lower than in MTLE without psychiatric comorbidities. Our results provide evidence of alterations in expression of NTs receptors in the epileptogenic hippocampus that are differentially modulated in presence of psychiatric comorbidities. As already explored in animal models, even in chronic human MTLE increased TrkB expression, among other NT receptors alterations, may play a major role in seizure type, frequency and surgery outcome.

Dihydrotestosterone deteriorates cardiac insulin signaling and glucose transport in the rat model of polycystic ovary syndrome

It is supposed that women with polycystic ovary syndrome (PCOS) are prone to develop cardiovascular disease as a consequence of multiple risk factors that are mostly related to the state of insulin resistance and consequent hyperinsulinemia. In the present study, we evaluated insulin signaling and glucose transporters (GLUT) in cardiac cells of dihydrotestosterone (DHT) treated female rats as an animal model of PCOS. Expression of proteins involved in cardiac insulin signaling pathways and glucose transporters, as well as their phosphorylation or intracellular localization were studied by Western blot analysis in DHT-treated and control rats. Treatment with DHT resulted in increased body mass, absolute mass of the heart, elevated plasma insulin concentration, dyslipidemia and insulin resistance. At the molecular level, DHT treatment did not change protein expression of cardiac insulin receptor and insulin receptor substrate 1, while phosphorylation of the substrate at serine 307 was increased. Unexpectedly, although expression of downstream Akt kinase and its phosphorylation at threonine 308 were not altered, phosphorylation of Akt at serine 473 was increased in the heart of DHT-treated rats. In contrast, expression and phosphorylation of extracellular signal regulated kinases 1/2 were decreased. Plasma membrane contents of GLUT1 and GLUT4 were decreased, as well as the expression of GLUT4 in cardiac cells at the end of androgen treatment. The obtained results provide evidence for alterations in expression and especially in functional characteristics of insulin signaling molecules and glucose transporters in the heart of DHT-treated rats with PCOS, indicating impaired cardiac insulin action.

Neurotrophin receptors expression in mesial temporal lobe epilepsy with and without psychiatric comorbidities and their relation with seizure type and surgical outcome

Epilepsy and psychiatric comorbidities are frequently associated, but their common biological substrate is unknown. We have previously reported altered structural elements and neurotrophins (NTs) expression in mesial temporal lobe epilepsy (MTLE) patients with psychiatric comorbidities. NTs receptors can regulate neurotransmission and promote neuroplasticity, being important candidates in the regulation and manifestation of psychopatological states and seizure-related events. MTLE hippocampi of subjects without psychiatric history, MTLE + major depression, MTLE + interictal psychosis derived from epilepsy surgery, and control necropsies were investigated for p75NTR, TrkB, TrkA, and TrkC immunohistochemistry. Increased expression of p75NTR, decreased TrkA, unaltered TrkC, and complex alterations involving TrkB expression were seen in MTLE groups. Increased TrkB expression in patients without complete seizure remission and in those with secondarily generalized seizures was seen. Decreased p75NTR expression associated with interictal psychosis, and increased TrkB in those with psychosis or major depression was also reported, although their p75NTR/TrkB ratios were lower than in MTLE without psychiatric comorbidities. Our results provide evidence of alterations in expression of NTs receptors in the epileptogenic hippocampus that are differentially modulated in presence of psychiatric comorbidities. As already explored in animal models, even in chronic human MTLE increased TrkB expression, among other NT receptors alterations, may play a major role in seizure type, frequency and surgery outcome.