• T Cell Factor-1 (TCF-1) Regulates Mature Alloactivated T Cells to Separate GVHD From GVL

      Mobin Karimi; Harris, Rebecca (2021)
      Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a curative treatment used for patients with cancer or other hematological malignancies. However, widespread use of this treatment is hindered by development of graft-versus-host disease (GVHD), a life-threatening complication of allo-HSCT. Mature donor T cells in the graft mediate GVHD, but also help kill residual malignant cells in the patient by the graft-versus-leukemia (GVL) effect. Depletion of mature T cells from the graft eliminates this beneficial anti-tumor response. Mature T cells are also needed for proper stem cell engraftment. Therefore, current work has focused on how to modulate T cell signaling and function to separate GVHD from GVL. T Cell Factor-1 (TCF-1) is a T cell developmental transcription factor that is also important in some contexts for T cell activation. The role of TCF-1 in alloactivated mature T cells is completely unknown. To examine the role of TCF-1 in this context, a mouse model of allo-HSCT leading to GVHD/GVL was used to study T cells from mice with a T cell-specific deletion of TCF-1. This work showed that loss of TCF-1 separates GVHD from GVL, with reduced disease severity and persistence yet maintained GVL effects. TCF-1 affects alloactivated T cell phenotypes and suppressive profiles, as well as the major T cell functions (proliferation, migration, and cytokine/cytotoxic mediator production). TCF-1 also controls alloactivated T cell survival, apoptosis, and gene expression programs. The regulation of these functions and programs by TCF-1 is distinct in CD4 versus CD8 T cells. TCF-1 also controls two unique T cell subsets - stem-like CXCR5+ CD8+ T cells, and CD25- noncanonical Tregs. Therefore, TCF-1, or these two unique T cell types, may be a therapeutic target to improve allo-HSCT outcomes by separating GVHD from GVL effects. Expansion of CD25- Tregs during TCF-1 deficiency may also be useful for treatment of other T cell-mediated disorders as well.
    • Targetingof PIM1 KinaseinMyeloproliferative NeoplasmsInduced by JAK2V617F

      Mohi, Golam; Stuver, Matthew (2017)
      Myeloproliferative neoplasms (MPNs) arestem cell-derivedblood disorders. The most common mutation found in MPN patients is the JAK2V617Fmutation. JAK2 is anon-receptor tyrosine kinase involved in STAT signaling. The JAK2V617F mutation is asingle amino acid substitution of a phenylalanine for valine, whichcauses JAK2 to be constitutively activated. This mutation can cause ahematopoietic transformation. Eventuallythis transformationcan lead to the development of one of thethree different Philadelphia-negative MPN diseases: Polycythemia Vera (PV), Essential Thrombocythemia (ET), and Primary Myelofibrosis (PMF). The JAK2V617F mutationhas been identified in 95% PVpatients,and 50-60% ofETand PMF patients.A JAK1/2 inhibitor (ruxolitinib) has been approved for MF and PV patients and,though it provides initial benefits, it is not effective enough to causelong-termremission in patients. This creates a critical need to identify new therapeutic targets for MPN patients. We found that PIM1 levels were significantly increased inMPN patients, as well asour JAK2V617F mouse modelof MPN.We observedthatknockdown of PIM1 caused a significant decrease in proliferationof JAK2V617F expressing cells. We also found that PIM1 knockdownhad no effect on the proliferation of hematopoietic cells not expressing JAK2V617F, leading us to believe PIM1 is only required in JAK2V617F mediated proliferation. Pharmacological inhibition of PIM kinases,using TP-3654,(kindly provided by Tolero pharmaceuticals)also led to a significant decrease in proliferation of JAK2V617F-expressing cells, but had no effect on cellslacking the mutation. We also found thatthePIM inhibitor,TP-3654,workssynergistically with ruxolitinibto achieve an even greater decrease in proliferation. We found that using the combination of ruxolitiniband TP-3654,we could use both drugs at lower concentrations andachieve an even greater decrease in proliferation and an increase apoptosis. Furthermore,we found that inhibition of PIM kinasesusing TP-3654can resensitize ruxolitinib-resistant cells to ruxolitinibtreatment.These important findingsshow that PIM1 plays animportantrolein the proliferation of hematopoietic cells expressing the JAK2V617F mutation, but is dispensable for the maintenance of cells lacking the mutation. We also found that targeting PIM kinases with TP-3654,significantly decreasedthe proliferation, and increaseapoptosisactivationof JAK2V617Fexpressing cells. We also showedthat TP-3654 and ruxolitinibcan work synergistically. Lastly, we showed that inhibition of PIM kinases,using TP-3654,caused ruxolitinib-resistant cells tobecome resensitized toruxolitinib. These findings helpedus come to the conclusionthatPIM1 kinase, is an importanttherapeutictargetin JAK2V617F-induced MPNs.
    • Teaching of SQL Through a Game

      Ward, Patrick T. (2015-05-01)
      The project seeks to provide an effective alternate method for teaching SQL through the use of a Game. There is value in learning SQL, as SQL skills are still in the top ten list of sought after IT skills for 2015 (Greenspan, 2014). However, lecture based teaching may not fully engage the learner. Therefore, the game was constructed with the MDA framework and Problem-Based Learning in mind. These methods help the learner bridge the gaps between lesson content, problems, and solutions. The game itself was constructed with MSSQL, Adobe Cold Fusion, HTML, CSS, and JavaScript. The game presents lessons on SQL concepts and quiz-based challenges for students to solve. Students faced increasingly difficult challenges as their level SQL knowledge expanded. The project may be found here: http://www.patrickward.net/SQLGame/
    • Technology Case Study in Storage Area Networks

      Marsh, John ; Thesis Advisor; Hash, Larry J.; Climek, David; Bull, Ronny; Pethe, Ameya (2014-05)
      In today's world we need immediate access to data. The demand for networked data access has increased exponentially in the last 20 years. With that demand the importance and volume of networked data has also grown exponentially. The speed at which the data can be accessed has increased and with that the data has moved from individual workstations to a networked location. Over the last decade there has been a trend to move mission critical data away from individual workstations to a centralized data center. A centralized data center removes the location constraint for accessing the data. If critical data is stored on individual servers, a failure will cause the data to be inaccessible. Today, mission critical applications are spanned over multiple servers for redundancy. With this topology, having the data in a central location allows the individual servers to better work with data. With the addition of virtualization, servers can be moved online from one physical server to another. If the data is centralized, it can be presented to all hosts in the cluster. This allows servers to move efficiently between hosts without losing access to the critical data. Many businesses in various industries like finance, airline, hospital, research, etc. depend on the speed and secure availability of their centralized data to function efficiently.
    • A Technology Case Study on Integrating Open Stack with SDN for Internet Connectivity using BGP

      Gonuguntla, Raja Bhushan Rao; Hash, Larry; Advisor (2016-12)
      There were many developments in Internet usage, which resulted in significant increase in Internet routing. With existing networking infrastructure, it is difficult to meet these requirements and causing more cost and less performance. Since network devices are hardware modules, processing them requires more power and more memory. However, if network protocols are developed using software modules, flexibility can be achieved in various programming applications and reduces dependency on hardware. The concept of using networking protocols as a software module can be explained using “Software Defined Networking (SDN).” With SDN, existing infrastructure can be integrated with various applications and centralized control protocols can be developed. One of the key components of SDN is integrating with Cloud Computing, where many applications can be built, which can be used for on-demand services. Integrating cloud computing with SDN will create dynamic networks and reduces infrastructure costs. In this paper, a case was considered for providing better internet connectivity by building public & private networks using Open source cloud technology (OpenStack) and existing distribution environments. For connectivity, BGP was used as routing protocol as it is known to be well- suited for such environments. Both public and private networks were integrated with SDN for centralized control. OpenStack was used to build various network topologies using different plugins through SDN controller. This method allowed to develop SDN controller with global view of OpenStack networks. The same controller was connected to distributed layers using Open Flow protocol. Since, both OpenStack and distributed networks were attached to SDN controller, centralized control of network protocols could be achieved. This model of centralized networks could be very useful in reducing costs and improving network efficiency, especially in large scale deployments.
    • Technology Case Study on Web Real-Time Communications (WebRTC)

      Karnati, Nagarjuna; Hash, Larry; Adviser (2016-05-01)
      Web real-time communication (WebRTC) is the latest technology standard which enables web browsers to communicate directly without having to install any internal or external plug-ins. WebRTC fills a critical gap in the web platform where a native proprietary app like Skype could do something which is media communication that World Wide Web just couldn’t. Now, this can be done form web using WebRTC technology. This paper starts with a brief introduction of WebRTC and how it got started. Moving on, it provides information about the WebRTC technical goals, architecture and protocols involved. This paper highlights the network address translation (NAT) traversal where STUN, TURN and ICE protocols are involved. Also, this paper highlights about the peer to peer to media flows with reference to WebRTC protocol stack and application program interface (API). In the end, this paper discusses about implemented security features, tools available for WebRTC development and provides enterprise use cases.
    • Text Detection from an Image

      Andriamanalimanana, Bruno R.; Thesis Advisor; Novillo, Jorge; Thesis Committee; Spetka, Scott; Thesis Committee; Goda, Piyush Jain (2020-12)
      Recently, a variety of real-world applications have triggered a huge demand for techniques that can extract textual information from images and videos. Therefore, image text detection and recognition have become active research topics in computer vision. The current trend in object detection and localization is to learn predictions with high capacity deep neural networks trained on a very large amount of annotated data and using a high amount of processing power. In this project, I have built an approach for text detection using the object detection technique. Our approach is to deal with the text as objects. We use an object detection method, YOLO (You Only Look Once), to detect the text in the images. We frame object detection as a regression problem to spatially separated bounding boxes and associated class probabilities. YOLO, a single neural network, that predicts bounding boxes and class probabilities directly from full images in one evaluation. Since the whole detection pipeline is a single network, it can be optimized end-to-end directly on detection performance. The MobileNet pre-trained deep learning model architecture was used and modified in different ways to find the best performing model. The goal is to achieve high accuracy in text spotting. Experiments on standard datasets ICDAR 2015 demonstrate that the proposed algorithm significantly outperforms methods in terms of both accuracy and efficiency.
    • The Effect of Multifocal Contact Lenses on Accomodation and Phoria in a Pediatric Population

      Gong, Celia (2017)
      The increasing prevalence of the use of distance-centered multifocal (MF) contact lenses as a method of myopia control in the pediatric population calls for a better understanding of binocularity and accommodation in children wearing these lenses. This was a prospective, randomized, crossover, single visit study that enrolled myopic children with normal accommodation and binocular vision and no history of myopia control treatment. All subjects were fitted with Coopervision Biofinity single vision (SV) and MF (+2.50D center distance add) contact lenses. Accommodative responses (photorefraction) and phorias (Modified Thorington) were measured at 4 distances (>3m, 100cm, 40cm, 25cm). Secondary measures included high and low contrast logMAR acuity, accommodative amplitude, and accommodative facility. Differences between MF and SV contact lenses were analyzed using repeated measures regression and paired t-tests. A total of 16 subjects, aged 10-15 years, completed the study. There was a small decrease in high (SV: -0.08, MF: +0.01) and low illumination (SV:-0.03, MF: +0.08) (both p<0.01) visual acuity, and contrast sensitivity (SV: 2.0 log units, MF: 1.9, p=0.015) with MFs. Subjects were more exophoric at 40cm (SV: -0.41 Δ, MF: -2.06 Δ) and 25cm (SV: -0.83 Δ, MF: -4.30 Δ) (both p<0.01). With MFs, subjects had decreased accommodative responses at distance (SV: -0.04 D; MF: -0.37 D, p=0.02), 100 cm (SV: +0.37 D; MF: -0.35 D, p<0.01), 40 cm (SV: +1.82 D; MF: +0.62 D, p<0.01), and 25 cm (SV: +3.38 D; MF: +1.75 D, p<0.01). There were no significant differences in accommodative amplitude (p=0.66) or accommodative facility (p=0.54). Children wearing MF contact lenses exhibited reduced accommodative responses and more exophoria at increasingly higher accommodative demands than with SV contact lenses. This suggests that children may be relaxing their accommodation and using the positive addition or increased depth of focus from added spherical aberration of the MF lenses. Further studies are needed to evaluate other lens designs, different amounts of positive addition and aberrations, and long-term adaptation to lenses.
    • THE ROLE OF DENGUE VIRUS NON-STRUCTURAL PROTEIN 1 IN DISEASE PATHOGENESIS

      King, Christine; Endy, Timothy; Barbachano-Guerrero, Arturo (2020)
      Dengue virus (DENV) causes an estimated 390 million infections worldwide annually, with severe forms of disease marked by vascular leakage and an over reactive inflammatory response. Endothelial cells (EC) are directly responsible for vascular homeostasis and are highly responsive to circulating mediators but are not commonly infected. Mast cells (MC) are potent cells of the innate immune system that play an important role in EC biology and inflammatory responses. DENV encodes 10 proteins; with only one, the non-structural protein 1 (NS1), secreted from infected cells and accumulating in the blood of patients.NS1 has been implicated in the pathogenesis of vascular permeability, but the mechanism is not completely understood. Using a complementary array of in vitroassays and disease relevant ECs and MCs, we described the possible roles for NS1 in dengue disease pathogenesis. Using microscopy and immunoblotting we observed that ECs internalize NS1 into endosomes, where it accumulates and is degraded overtime. Transcriptome and pathway analysis defined changes in global gene expression in ECs that are associated with cell dysfunction. We observed that NS1 induced an increase in multicellular rearrangements and a decrease in barrier function in ECs. We demonstrated that NS1-dependent activation of the p38 MAPK pathway controls the changes in EC permeability in vitro. Further, we discovered iiithat ECs and MCs respond to NS1 by secreting a specific array of proinflammatory cytokines and chemokines that may contribute to the cytokine storm in dengue disease. Finally, we found that NS1 internalization can mediate the uptake of bound antibodies into ECs. Together, these results suggest a vasoactive and proinflammatory role for DENV NS1 that may participate in the development of severe symptoms in dengue disease. The observed functions of NS1 could lead to the discovery of new therapeutic targets in dengue disease.
    • The Ties That Bind: A blog project about the meaning of fandom

      Stam, Kathryn; Thesis Advisor; Lizardi, Ryan; Second Reader; Isgar, Eric (2021-05)
      Fandoms, or fan communities; groups of enthusiasts or ‘fans’ that have come together through their shared love of some kind of media, have been around for a long time in a number of different forms around the world, though in small numbers. When comic books became popular, those groups increased, and once again as movies became more commonplace. These communities and cultures are reliant on information technology, and the technology reliant on them, in a symbiotic relationship. It is my objective to research, analyze and observe fandoms and the culture related to the associated communities of fans within, and how they have formed, communicated and interacted, both prior to the development of modern information technology and after, as well as their continued growth on established and upcoming platforms. My actual project consists of a set of blog posts, or ‘mini-essays’, on Tumblr, around one hundred words per post, perhaps more, with ten posts in total.
    • TIMP-2 PHOSPHORYLATION BY EXTRACELLULAR c-SRC REGULATES proMMP-2 ACTIVATION

      Bourboulia, Dimitra; Sánchez Pozo, Javier (2018)
      Matrix metalloproteinases (MMPs) are secreted zinc-dependent endopeptidases that are involved in many extracellular biological processes due to their matrix-degrading function. The majority of theseenzymes are released intothe extracellular space in their inactive form and require activation. The tissue inhibitors of metalloproteinases (TIMPs) are also secreted proteins and mainly function to inhibit all members of the MMP family. Interestingly, TIMP-2 also participates in the activationprocessof proMMP-2. Although the interaction between TIMP-2 and proMMP-2 has been known for decades, the molecular signal that triggersthis association has only recently beendetermined. Studies in our lab haveshown that TIMP-2 is tyrosine phosphorylatedby the c-Srctyrosinekinase. Also, phosphorylation of TIMP-2 Tyr90is essential for its interaction with proMMP-2in vivo. Our hypothesis is that c-Src-mediated TIMP-2 phosphorylation happens outside the cell. Here, wedemonstratethat TIMP-2 and c-Src are secreted through different secretory pathways and that TIMP-2 phosphorylation takes place in the extracellular space. Our workalso showsthatextracellularc-Srcisactive, reinforcing the fact that phosphorylation can happen extracellularly. We also hypothesize that extracellular c-Src plays a critical role in facilitating TIMP-2:proMMP-2 interaction. We first confirmed thatTIMP-2 and proMMP-2 endogenously interact only in cells containing endogenous c-Src. This interaction,as well as TIMP-2 phosphorylation,was blocked by treating cells with acustom-made anti-c-Src polyclonal antibody (pAb)that targets amino acids 84-110. We also showthat ananti-c-Src antibody that targets the first 79 amino acids does not inhibit TIMP-2 phosphorylation and interaction with proMMP-2. Therefore, since TIMP-2:proMMP-2 complex formation promotes proMMP-2 activation, we hypothesize that c-Src is an essential player in this process. Our data showsthatthe non-phosphorylatable TIMP-2Tyr90mutant does not promote proMMP-2 activation. Furthermore, pretreatment with the anti-c-Src pAbblockedTIMP-2-mediated proMMP-2 activation, whereasthe anti-c-Src mAb6 did not affect proMMP-2 activation. Overall, these findings provide further evidence that secreted c-Src-mediated TIMP-2 phosphorylation occurs in the extracellular space, where thesecretedkinase is also active. Moreover, c-Src is essential for TIMP-2:proMMP-2 complex formation as well as proMMP-2 activation.
    • To Upgrade or Not To Upgrade Application

      Stam, Kathryn; Thesis Advisor; Lizardi, Ryan; Second Reader; Francisco, Neil (2021-05)
      New Technology consists of new hardware devices, computational workflows, digital advances, and information systems. As technology continues to evolve over the years, this never-ending cycle of new devices and experiences will always be present amongst consumers. Traditionally, new hardware devices are intriguing because they are designed to improve our access to information, media, and a connection to the digital world, but does this mean our previous-gen devices are no longer valuable? This project involves creating a prototype application designed for both computer and mobile interfaces to help improve the accessibility to information and the overall user experience with an older device. The “To Upgrade or Not To Upgrade” app will inform end-users of their older technological device specifications and suggest hardware/software methods to unlock their full potential. The goal of this paper is to shed some light on consumers that upgrading to the following gen devices is not always necessary to receive the best human-to-computer interactions. It is likely the computer or mobile device that one owns now, with some slight modifications, is all that is needed to provide a pleasant experience.
    • A trancriptomics based approach reveals the functional consequences of RNase MRP RNA mutations in yeast.

      Schmitt, Mark; Shafiuddin, Md (2018)
      RNase MRP is a eukaryotic ribonucleoprotein complex involved in multiple cellular functions that includes ribosomal RNA processing, primer generation for mitochondrial DNA replication and degradation of cell cycle related mRNAs. In Saccharomyces cerevisiae, the RNA component of RNase MRP is encoded by NME1. We have performed random deletion mutagenesis of RNase MRP RNA gene and isolated a mutation, nme1-91, that causes temperature sensitive growth defect on glycerol media. RNA analysis of nme1-91 showed that this mutant is mildly deficient in the 5.8S rRNA processing function of RNase MRP. Growth analysis and northern blotting of RNase MRP RNA mutations generated based on nme1-91 allele suggested that 3’-end nucleotide sequences of the nme1-91 allele contribute to its phenotype. Highcopy suppression screen identified tRNA modification gene NCS6 as a suppressor of nme1-91. Additionally, primary mode of suppression by NCS6 was found to be non-mitochondrial since NCS6 partially suppressed the nme1-91 phenotype on fermentable carbon source. Strains carrying a deletion of NCS6 in combination with nme1-91 showed a synthetic sick phenotype. Polysome profile analysis of nme1-91 revealed that 80S monosomal fraction accumulates in this mutant. Differential gene expression analysis of nme1-91 by RNAseq indicated that rRNA processing and cell cycle related genes become mis-regulated due to this mutation. A similar high-throughput sequencing based approach was also employed to investigate the transcriptional basis of positive genetic interactions between components of RNase MRP and nonsense-mediated decay pathway. A yeast strain bearing the nme1-P6 mutation in the RNA component of RNase MRP exhibits temperature-sensitive growth defect. This phenotype can be suppressed by deletion of NMD components. Differential gene expression analysis identified several mis-regulated biological processes in nme1-P6 and Δupf1 strains. Comparative transcriptomic analysis suggested that suppression of nme1-P6 phenotype by Δupf1 is accompanied by large shift in gene expression pattern towards Δupf1 strain. Moreover, the majority of direct targets of NMD were not down-regulated in nme1-P6 indicating that the effect of NMD on nme1-P6 might be due to increased degradation on mRNAs that are not targeted by NMD in normal conditions. Taken together, these results show that mutations of RNase MRP RNA can modulate diverse biological processes.
    • Transcriptome-wide gene expression in a rat model of attention deficit hyperactivity disorder symptoms: Rats developmentally exposed to polychlorinated biphenyls

      Sazonova, Nadezhda A.; DasBanerjee, Tania; Middleton, Frank A.; Gowtham, Sriharsha; Schuckers, Stephanie; Faraone, Stephen V. (Wiley, 2011-09-14)
      Polychlorinated biphenyls (PCB) exposure in rodents provides a useful model for the symptoms of Attention deficit hyperactivity disorder (ADHD). The goal of this study is to identify genes whose expression levels are altered in response to PCB exposure. The brains from 48 rats separated into two age groups of 24 animals each (4 males and 4 females for each PCB exposure level (control, PCB utero, and PCB lactational)) were harvested at postnatal days 23 and 35, respectively. The RNA was isolated from three brain regions of interest and was analyzed for differences in expression of a set of 27,342 transcripts. Two hundred seventy-nine transcripts showed significant differential expression due to PCB exposure mostly due to the difference between PCB lactational and control groups. The cluster analysis applied to these transcripts revealed that significant changes in gene expression levels in PFC area due to PCB lactational exposure. Our pathway analyses implicated 27 significant canonical pathways and 38 significant functional pathways. Our transcriptomewide analysis of the effects of PCB exposure shows that the expression of many genes is dysregulated by lactational PCB exposure, but not gestational exposure and has highlighted biological pathways that might mediate the effects of PCB exposure on ADHD-like behaviors seen in exposed animals. Our work should further motivate studies of fatty acids in ADHD, and further suggests that another potentially druggable pathway, oxidative stress,may play a role in PCB inducedADHD behaviors
    • Traumatic Brain Injury and Schizophrenia in Members of Schizophrenia and Bipolar Disorder Pedigrees

      Malaspina, Dolores; Goetz, Raymond R.; Friedman, Jill Harkavy; Kaufmann, Charles A.; Faraone, Stephen V.; Tsuang, Ming; Cloninger, C. Robert; Nurnberger, John I.; Blehar, Mary C. (American Psychiatric Association Publishing, 2001-03)
      Objective: Schizophrenia following a traumatic brain injury could be a phenocopy of genetic schizophrenia or the consequence of a gene-environment interaction. Alternatively, traumatic brain injury and schizophrenia could be spuriously associated if those who are predisposed to develop schizophrenia have greater amounts of trauma for other reasons. The authors investigated the relationship between traumatic brain injury and psychiatric diagnoses in a large group of subjects from families with at least two biologically related first-degree relatives with schizophrenia, schizoaffective disorder, or bipolar disorder. Method: The Diagnostic Interview for Genetic Studies was used to determine history of traumatic brain injury and diagnosis for 1,275 members of multiplex bipolar disorder pedigrees and 565 members of multiplex schizophrenia pedigrees. Results: Rates of traumatic brain injury were significantly higher for those with a diagnosis of schizophrenia, bipolar disorder, and depression than for those with no mental illness. However, multivariate analysis of within-pedigree data showed that mental illness was related to traumatic brain injury only in the schizophrenia pedigrees. Independent of diagnoses, family members of those with schizophrenia were more likely to have had traumatic brain injury than were members of the bipolar disorder pedigrees. The members of the schizophrenia pedigrees also failed to show the gender difference for traumatic brain injury (more common in men than in women) that was expected and was present in the bipolar disorder pedigrees. Subjects with a schizophrenia diagnosis who were members of the bipolar disorder pedigrees (and thus had less genetic vulnerability to schizophrenia) were less likely to have had traumatic brain injury (4.5%) than were subjects with schizophrenia who were members of the schizophrenia pedigrees (and who had greater genetic vulnerability to schizophrenia) (19.6%). Conclusions: Members of the schizophrenia pedigrees, even those without a schizophrenia diagnosis, had greater exposure to traumatic brain injury compared to members of the bipolar disorder pedigrees. Within the schizophrenia pedigrees, traumatic brain injury was associated with a greater risk of schizophrenia, consistent with synergistic effects between genetic vulnerability for schizophrenia and traumatic brain injury. Posttraumatic-braininjury schizophrenia in multiplex schizophrenia pedigrees does not appear to be a phenocopy of the genetic disorder.
    • Treatment of nonpsychotic relatives of patients with schizophrenia: Six case studies

      Tsuang, Ming T.; Stone, William S.; Tarbox, Sarah I.; Faraone, Stephen V. (Wiley, 2002-11-27)
      There is growing support for the notion that the genetic liability for schizophrenia could be manifested in brain dysfunction, even without the full manifestations of schizophrenia [Meehl, 1962, 1989; Seidman, 1997; Faraone et al., 2001]. This liability is characterized clinically by neurologic, neurobiological, psychiatric, neuropsychological, and psychosocial impairments in nonpsychotic, first-degree relatives of people with schizophrenia and includes eye tracking dysfunction [Levy et al., 1994], allusive thinking [Catts et al., 1993], neurologic signs [Erlenmeyer-Kimling et al., 1982], biochemical abnormalities [Callicott et al., 1998], char acteristic auditory evoked potentials [Friedman and Squires-Wheeler, 1994], neuroimaging assessed brain abnormalities [Seidman et al., 1997], and neuropsycho logical impairment [Kremen et al., 1994]. Paul Meehl introduced the term ‘‘schizotaxia’’ in 1962 to describe the genetic predisposition to schizophrenia [Meehl, 1962], and we have modified the concept to take account of subsequent research [Faraone et al., 2000]. The concept of schizotaxia raises at least three fundamental issues: 1) What is the conceptual basis of schizotaxia? 2) Is it a valid syndrome? and 3) perhaps most importantly from the point of view of the eventual prevention of schizo phrenia, is it treatable? In this paper, we review the model of schizotaxia by focusing first on its nature and extent. We then describe preliminary research criteria for its diagnosis in nonpsychotic relatives of schizo phrenic patients, followed by a presentation of our initial attempts to treat schizotaxia. Finally, prospects for the future focus on the need to validate the proposed syndrome further and on the clinical implications of treating schizotaxia.
    • Triptolide Induces Increases in Migration via Mitogen-activated Protein Kinase Phosphatase-1 control of P38 and JNK MAPK Activation

      Parekh, Nili M. (2010-07-16)
      Purpose Triptolide is a Chinese herbal extract known for its anti-inflammatory and immunosuppressive effects in treating chronic inflammatory diseases and tumors. As these attributes promote wound healing, we determined if triptolide enhances corneal wound healing by stimulating human corneal epithelial cell (HCEC) migration through changes in negative feedback regulation by a dual specificity protein Phosphatase (DUSP1) of MAPK signaling mediated effect. Methods SV40-adenovirus-immortalized HCEC were maintained in DMEM/F12. Specific shRNA for MKP-1(DUSP1) and c-jun NH2- terminal kinase JNK-1 were transduced to establish stable cell lines deficient in their respective gene expression. Scratch wound assay was employed to assess cell migration rates by taking time-dependent serial photographs of cells following wound creation. Hydroxyurea (2.5 mM) was also added to the medium to inhibit cell proliferation during the experiment. Cell Titer-Glo® luminescent cell viability assay was used to evaluate cell viability by measuring ATP production by HCEC. Results Triptolide did not affect cell viability up to 10nM and stimulated wound closure through increases in migration. Maximal responses occurred at 1nM. These increases in migration were suppressed below that in the untreated control when p38 or JNK MAPK activation was inhibited. In the MKP-1 knockdown cells, migration was stimulated relative to the control and triptolide failed to augment this response. In JNK-1 knockdown cells, migration is comparable to SV40 wild type cells. In JNK-1 knockdown cells, triptolide mediated increases are diminished completely in the presence of p38 inhibition. Conclusions Triptolide at concentrations up to 10 nM promotes cell migration without compromising cell survival. Such promotion is mediated by loss of MKP-1 negative feedback control of p38 and JNK activation. Therefore, triptolide stimulates cell migration through inhibition of MKP-1 (DUSP1) stabilization induced by kinase mediated phosphorylation.
    • THE TUMOR SUPPRESSIVE ROLE OF MONOGLYCERIDE LIPASE IN LUNG CANCER

      Huang, Ying; Liu, Renyan (2017)
      Monoglyceride lipase (MGL) is a serine hydrolase that hydrolyzes 2-monoglycerides and produces fatty acid and glycerol. Our previous studies showed that there was MGL deficiency in the majority of human lung, breast, and colorectal cancer tissues as compared with normal tissues. Further studies suggested that MGL was a potential tumor-suppressor in the development of colorectal cancer. However, paradoxical findings about the role of MGL in tumorigenesis have been reported. It is therefore important to further elucidate the function of MGL in tumorigenesis. To that end, we generated MGL-knockout mouse and found that MGL knockout led to tumor formation in multiple organs/tissues of mice. Particularly, the major findings were lung adenocarcinomas. In cultured cells, MGL deletion enhanced cell proliferation and induced cellular transformation. Further molecular studies demonstrated that MGL suppressed EGFR signaling, NF-κB activity, and COX-2 expression. Deficiency of MGL may lead to over-activation of EGFR, NF-κB, and COX-2, and therefore contribute to tumorigenesis. We also found that MGL over-expression induced remarkable cancer cell apoptosis, and increased cleavage of caspase-8, caspase-9, caspase-3 and PARP. MGLinduced apoptosis thus involves both the intrinsic mitochondria-mediated and extrinsic death receptor-initiated apoptosis pathways. Most importantly, our data indicated that MGL interacted with a potent inhibitor of apoptosis, XIAP, and significantly reduced XIAP protein stability, which may unleash caspase-9 and caspase-3 from XIAP inhibition and thereby promote apoptosis. Some additional findings about MGL showed that MGL localization to lipid droplets was important for its regulation of cell growth and pro-tumorigenic signaling while MGL’s lipase activity is dispensable for these effects. We identified Asp-115 (D115) of MGL as the most important amino acid residue in mediating MGL localization to lipid droplets. We also found that MGL deficiency promotes lipid droplet formation and cellular lipid accumulation, which potentially promotes cancer cell survival under stressful conditions. Overall, our in vitro and in vivo data strongly support the notion that MGL is a tumor suppressor. The molecular findings about MGL may have revealed certain targets for personalized cancer therapy in the context of MGL deficiency and they also implicate potential roles of MGL in inflammation, immunity, and metabolism.
    • THE TUMOR SUPPRESSOR FOLLICULIN REGULATES GLYCOLYSIS BY SPECIFICALLY BINDING AND INHIBITING LACTATE DEHYDROGENASE-A

      Mollapour, Mehdi; Woodford, Mark R. (2021)
      Folliculin (FLCN) is tumor suppressor protein whose function remains a topic of debate. Germline mutations in FLCN predispose affected individuals to develop Birt-Hogg-Dubé syndrome, which is characterized by facial fibrofolliculomas, pulmonary cysts, spontaneous pneumothorax, and renal cell carcinoma. These kidney tumors exhibit an elevated glycolytic phenotype even in the presence of oxygen, an observation commonly known as the “Warburg effect.” This phenomenon is driven by the activity of lactate dehydrogenase-A (LDHA), the enzyme responsible for the interconversion of pyruvate and lactate in the terminal step of glycolysis. Our work herein shows that FLCN is a specific intracellular inhibitor of LDHA. Biochemical and biophysical analyses mapped the interaction and inhibition of LDHA activity to an unstructured loop in FLCN positioned between the amino and carboxy-terminal domains. Characterization of a minimal 10-amino acid FLCN-derived peptide demonstrated that it was sufficient to both bind and inhibit LDHA activity in vitro. Further, treatment of FLCN-deficient cells with this FLCN-derived peptide is sufficient to suppress glycolysis. Interestingly, evaluated cancer cell lines derived from solid tumors of the lung, breast, prostate, bladder, and colon also demonstrate dysregulation of LDHA activity and dissociation of FLCN-LDHA interaction. Previous work has shown that inhibition of LDHA in cancer cells leads to apoptosis. Accordingly, treatment of these cell lines with the FLCN-peptide results in apoptosis, suggesting a potential therapeutic intervention. Taken together, inhibition of LDHA by the tumor suppressor FLCN provides a mechanistic explanation for the endogenous regulation of glycolysis.