Pathology Commons^™

Catabolic Degradation Of Endothelial Vegfa Via Autophagy, Thomas Neill, Carolyn Chen, Simone Buraschi, Renato V. Iozzo

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Extracellular matrix-evoked angiostasis and autophagy within the tumor microenvironment represent two critical, but unconnected, functions of the small leucine-rich proteoglycan, decorin. Acting as a partial agonist of vascular endothelial growth factor 2 (VEGFR2), soluble decorin signals via the energy sensing protein, AMP-activated protein kinase (AMPK), in the autophagic degradation of intracellular vascular endothelial growth factor A (VEGFA). Here, we discovered that soluble decorin evokes intracellular catabolism of endothelial VEGFA that is mechanistically independent of mTOR, but requires an autophagic regulator, paternally expressed gene 3 (PEG3). We found that administration of autophagic inhibitors such as chloroquine or bafilomycin A1, or depletion …

Pbrm1 Acts As A P53 Lysine-Acetylation Reader To Suppress Renal Tumor Growth., Weijia Cai, Liya Su, Lili Liao, Zongzhi Z Liu, Lauren Langbein, Essel Dulaimi, Joseph R Testa, Robert G Uzzo, Zhijiu Zhong, Wei Jiang, Qin Yan, Qing Zhang, Haifeng Yang

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

p53 acetylation is indispensable for its transcriptional activity and tumor suppressive function. However, the identity of reader protein(s) for p53 acetylation remains elusive. PBRM1, the second most highly mutated tumor suppressor gene in kidney cancer, encodes PBRM1. Here, we identify PBRM1 as a reader for p53 acetylation on lysine 382 (K382Ac) through its bromodomain 4 (BD4). Notably, mutations on key residues of BD4 disrupt recognition of p53 K382Ac. The mutation in BD4 also reduces p53 binding to promoters of target genes such as CDKN1A (p21). Consequently, the PBRM1 BD4 mutant fails to fully support p53 transcriptional activity and is defective …

Multiple Mitochondrial Thioesterases Have Distinct Tissue And Substrate Specificity And Coa Regulation, Suggesting Unique Functional Roles., Carmen Bekeova, Lauren Anderson-Pullinger, Kevin Boye, Felix Boos, Yana Sharpadskaya, Johannes M Herrmann, Erin L. Seifert

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Acyl-CoA thioesterases (Acots) hydrolyze fatty acyl-CoA esters. Acots in the mitochondrial matrix are poised to mitigate β-oxidation overload and maintain CoA availability. Several Acots associate with mitochondria, but whether they all localize to the matrix, are redundant, or have different roles is unresolved. Here, we compared the suborganellar localization, activity, expression, and regulation among mitochondrial Acots (Acot2, -7, -9, and -13) in mitochondria from multiple mouse tissues and from a model of Acot2 depletion. Acot7, -9, and -13 localized to the matrix, joining Acot2 that was previously shown to localize there. Mitochondria from heart, skeletal muscle, brown adipose tissue, and …

New Insights Into The Lactate Shuttle: Role Of Mct4 In The Modulation Of The Exercise Capacity., Sara Bisetto, Megan C Wright, Romana A Nowak, Angelo C Lepore, Tejvir S Khurana, Emanuele Loro, Nancy J Philp

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Lactate produced by muscle during high-intensity activity is an important end product of glycolysis that supports whole body metabolism. The lactate shuttle model suggested that lactate produced by glycolytic muscle fibers is utilized by oxidative fibers. MCT4 is a proton coupled monocarboxylate transporter preferentially expressed in glycolytic muscle fibers and facilitates the lactate efflux. Here we investigated the exercise capacity of mice with disrupted lactate shuttle due to global deletion of MCT4 (MCT4−/−) or muscle-specific deletion of the accessory protein Basigin (iMSBsg−/−). Although MCT4−/− and iMSBsg−/− mice have normal muscle morphology and contractility, only MCT4−/− mice exhibit an exercise intolerant …

A Physiologically-Based Pharmacokinetic Model For Targeting Calcitriol-Conjugated Quantum Dots To Inflammatory Breast Cancer Cells., James Forder, Mallory Smith, Margot Wagner, Rachel J. Schaefer, Jonathan Gorky, Kenneth L. Van Golen, Anja Nohe, Prasad Dhurjati

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Quantum dots (QDs) conjugated with 1,25 dihydroxyvitamin D3 (calcitriol) and Mucin-1 (MUC-1) antibodies (SM3) have been found to target inflammatory breast cancer (IBC) tumors and reduce proliferation, migration, and differentiation of these tumors in mice. A physiologically-based pharmacokinetic model has been constructed and optimized to match experimental data for multiple QDs: control QDs, QDs conjugated with calcitriol, and QDs conjugated with both calcitriol and SM3 MUC1 antibodies. The model predicts continuous QD concentration for key tissues in mice distinguished by IBC stage (healthy, early-stage, and late-stage). Experimental and clinical efforts in QD treatment of IBC can be augmented by in …

Case-Based Asynchronous Interactive Modules In Undergraduate Medical Education., Tatiana Villatoro, Katherine Lackritz, Joanna S Y Chan

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Undergraduate medical education traditionally consists of 2 years of lecture-based courses followed by 2 years of clinical clerkships. However, over the past couple decades, undergraduate medical education has been evolving toward non-lecture-based integrated curriculums, requiring a collaborative curriculum. Additionally, e-learning platforms have become efficacious and essential to delivering education asynchronously to students. At Thomas Jefferson University, the Pathology and Obstetrics and Gynecology departments collaborated to create a pilot series of case-based asynchronous interactive modules to teach gynecologic pathology in a clinical context, while interweaving other educational components, such as evidence-based medicine, clinical skills, and basic sciences. The case-based asynchronous interactive …

Dysregulation Of Mitochondrial Ca2+ Uptake And Sarcolemma Repair Underlie Muscle Weakness And Wasting In Patients And Mice Lacking Micu1, Valentina Debattisti, Adam Horn, Raghavendra Singh, Erin L. Seifert, Marshall W. Hogarth, Davi A. Mazala, Kai Ting Huang, Rita Horvath, Jyoti K. Jaiswal, György Hajnóczky

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Muscle function is regulated by Ca²⁺, which mediates excitation-contraction coupling, energy metabolism, adaptation to exercise, and sarcolemmal repair. Several of these actions rely on Ca²⁺ delivery to the mitochondrial matrix via the mitochondrial Ca²⁺ uniporter, the pore of which is formed by mitochondrial calcium uniporter (MCU). MCU's gatekeeping and cooperative activation are controlled by MICU1. Loss-of-protein mutation in MICU1 causes a neuromuscular disease. To determine the mechanisms underlying the muscle impairments, we used MICU1 patient cells and skeletal muscle-specific MICU1 knockout mice. Both these models show a lower threshold for MCU-mediated Ca²⁺ uptake. Lack of …

Perturbed Mitochondria-Er Contacts In Live Neurons That Model The Amyloid Pathology Of Alzheimer's Disease., Pamela V. Martino Adami, Zuzana Nichtova, David B. Weaver, Adam Bartok Dr., Thomas Wisniewski, Drew R. Jones, Sonia Do Carmo, Eduardo M. Castaño, A. Claudio Cuello, György Hajnóczky, Laura Morelli

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

The use of fixed fibroblasts from familial and sporadic Alzheimer's disease patients has previously indicated an upregulation of mitochondria-ER contacts (MERCs) as a hallmark of Alzheimer's disease. Despite its potential significance, the relevance of these results is limited because they were not extended to live neurons. Here we performed a dynamic in vivo analysis of MERCs in hippocampal neurons from McGill-R-Thy1-APP transgenic rats, a model of Alzheimer's disease-like amyloid pathology. Live FRET imaging of neurons from transgenic rats revealed perturbed 'lipid-MERCs' (gap width <10 nm), while 'Ca2+-MERCs' (10-20 nm gap width) were unchanged. In situ TEM showed no significant differences in the lipid-MERCs:total MERCs or lipid-MERCs:mitochondria ratios; however, the average length of lipid-MERCs was significantly decreased in neurons from transgenic rats as compared to controls. In accordance with FRET results, untargeted lipidomics showed significant decreases in levels of 12 lipids and bioenergetic analysis revealed respiratory dysfunction of mitochondria from transgenic rats. Thus, our results reveal changes in MERC structures coupled with impaired mitochondrial functions in Alzheimer's disease-related neurons.This article has an associated First Person interview with the first author of the paper.

Mindfulness Based Stress Reduction Increases Pathology Resident Self Wellness, Kaitlin Collura, Md, Phd, Joanna Chan, Md

Department of Pathology, Anatomy, and Cell Biology Posters

Stress during medical residency is a common, complex issue influenced by time demands, level of social support, and environmental factors. Increased stress levels can lead to resident burnout, and may contribute to the high rates of depression and suicide seen among physicians. In 2015, the ACGME began a campaign to promote resident wellness by calling on programs to implement strategies to develop resiliency, identify problems, and promote wellbeing. Mindfulness based stress reduction (MBSR) strategies have been shown to decrease burnout, improve mood, and increase compassion in healthcare providers. We implemented a wellness retreat to educate residents on the principles of …

Wellness Initiative Program And Effect On Pathology Resident Burnout Rate, Kaitlin Collura, Md, Phd, Michelle Nagurney, Md, Joanna Chan, Md

Department of Pathology, Anatomy, and Cell Biology Posters

Increasing rates of physician burnout are reported across many medical specialties, including Pathology. In 2014, 52.5% of pathologists experienced symptoms of burnout compared to 37.6% in 2011. Suicide and depression rates in physicians are also higher than the average population. As a result, the ACGME began a campaign, in 2015, to foster resident wellness. The campaign calls for strategies to develop resiliency, identify problems, and promote well-being. While wellness initiatives may not change underlying mental illness, they may mitigate environmental factors that can exacerbate mental illness or suicidal behavior. Here we report results from a recently implemented resident wellness initiative …

Conceptualization Of A Parasympathetic Endocrine System., Jonathan Gorky, James Schwaber

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

We here propose a parasympathetic endocrine system (PES) comprised of circulating peptides released from secretory cells in the gut, significantly modulated by vagal projections from the dorsal motor nucleus of the vagus (DMV). While most of these gut peptides mediate well-described satiety and digestive effects that increase parasympathetic control of digestion (Lee et al., 1994; Gutzwiller et al., 1999; Klok et al., 2007), they also have actions that are far-reaching and increase parasympathetic signaling broadly throughout the body. The actions beyond satiety that peptides like somatostatin, cholecystokinin, glucagon-like peptide 1, and vasoactive intestinal peptide have been well-examined, but not in …

Comparison Of Two Commercial Matrix-Assisted Laser Desorption/Ionization-Time Of Flight Mass Spectrometry (Maldi-Tof Ms) Systems For Identification Of Nontuberculous Mycobacteria., Barbara A. Brown-Elliott, Thomas R. Fritsche, Brooke J. Olson, Sruthi Vasireddy, Ravikiran Vasireddy, Elena Iakhiaeva, Diana Alame, Richard J. Wallace, John A. Branda

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Objectives: This multi-center study’s aim was to assess the performance of two commercially-available matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems in identifying a challenge collection of clinically-relevant nontuberculous mycobacteria (NTM).

Methods: NTM clinical isolates (N=244) belonging to 23 species/subspecies were identified by gene sequencing and analyzed using the Bruker Biotyper with Mycobacterial Library v5.0.0 and the bioMérieux VITEK MS with v3.0 database.

Results: Using the Bruker or bioMérieux systems, 92% or 95% of NTM strains, respectively, were identified at least to the complex/group level; 62% and 57%, respectively, were identified to the highest taxonomic level. Differentiation between members …

Incidence Rates Of Critical Low Glucoses (<40 Mg/Dl) By Poct Before And After New Policies For Treatment Of Clinically Significant Hypoglycemia (<54 Mg/Dl): A Comparison Between Two Hospitals, Lilah Evans, Douglas F. Stickle, Barbara Goldsmith

Department of Pathology, Anatomy, and Cell Biology Posters

Glucose <54 mg/dL was recently defined as Clinically Significant Hypoglycemia (CSH, American Diabetes Association, Standards of Medical Care in Diabetes, 2017). In January 2018, our hospitals instituted a callback policy for inpatient CSH from the central laboratory, and instituted a new nursing procedure for response to CSH. We examined whether the new policies had affected the incidence rate of low glucose critical values (CRITICAL, <40 mg/dL) among POCT glucose measurements. This was investigated for two hospitals within our system: A, a 950-bed academic medical center hospital, and B, a 200-bed community hospital.

Mdivi-1, A Mitochondrial Fission Inhibitor, Modulates T Helper Cells And Suppresses The Development Of Experimental Autoimmune Encephalomyelitis., Yan-Hua Li, Fang Xu, Rodolfo Thome, Min-Fang Guo, Man-Luan Sun, Guo-Bin Song, Rui-Lan Li, Zhi Chai, Bogoljub Ciric, A. M. Rostami, Mark T. Curtis, Cun-Gen Ma, Guang-Xian Zhang

Department of Neurology Faculty Papers

BACKGROUND: Unrestrained activation of Th1 and Th17 cells is associated with the pathogenesis of multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). While inactivation of dynamin-related protein 1 (Drp1), a GTPase that regulates mitochondrial fission, can reduce EAE severity by protecting myelin from demyelination, its effect on immune responses in EAE has not yet been studied.

METHODS: We investigated the effect of Mdivi-1, a small molecule inhibitor of Drp1, on EAE. Clinical scores, inflammation, demyelination and Drp1 activation in the central nervous system (CNS), and T cell responses in both CNS and periphery were determined.

RESULTS: Mdivi-1 effectively …

Spg7 Targets The M-Aaa Protease Complex To Process Mcu For Uniporter Assembly, Ca2 Influx, And Regulation Of Mitochondrial Permeability Transition Pore Opening, Stephen Hurst, Ariele Baggett, György Csordás, Shey-Shing Sheu

Center for Translational Medicine Faculty Papers

The mitochondrial matrix ATPase associated with diverse cellular activities (m-AAA) protease spastic paraplegia 7 (SPG7) has been recently implicated as either a negative or positive regulatory component of the mitochondrial permeability transition pore (mPTP) by two research groups. To address this controversy, we investigated possible mechanisms that explain the discrepancies between these two studies. We found that loss of the SPG7 gene increased resistance to Ca²-induced mPTP opening. However, this occurs independently of cyclophilin D (cyclosporine A insensitive) rather it is through decreased mitochondrial Ca² concentrations and subsequent adaptations mediated by impaired formation of functional mitochondrial Ca …

Gangliosides: Treatment Avenues In Neurodegenerative Disease., Pierre J. Magistretti, Fred H. Geisler, Jay S. Schneider, P. Andy Li, Hubert Fiumelli, Simonetta Sipione

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Gangliosides are cell membrane components, most abundantly in the central nervous system (CNS) where they exert among others neuro-protective and -restorative functions. Clinical development of ganglioside replacement therapy for several neurodegenerative diseases was impeded by the BSE crisis in Europe during the 1990s. Nowadays, gangliosides are produced bovine-free and new pre-clinical and clinical data justify a reevaluation of their therapeutic potential in neurodegenerative diseases. Clinical experience is greatest with monosialo-tetrahexosyl-ganglioside (GM1) in the treatment of stroke. Fourteen randomized controlled trials (RCTs) in overall >2,000 patients revealed no difference in survival, but consistently superior neurological outcomes vs. placebo. GM1 was shown …

Extreme Peripheral Blood Plasmacytosis Mimicking Plasma Cell Leukemia As A Presenting Feature Of Angioimmunoblastic T-Cell Lymphoma (Aitl)., Kelsey Sokol, Saritha Kartan, William T. Johnson, Onder Alpdogan, Neda Nikbakht, Bradley M. Haverkos, Jerald Z. Gong, Pierluigi Porcu

Department of Medical Oncology Faculty Papers

Angioimmunoblastic T-cell lymphoma (AITL) is one of four major subtypes of nodal peripheral T cell lymphoma, characterized by its cell of origin, the follicular helper T-cell (TFH). Patients typically present with prominent constitutional (B) symptoms, generalized lymphadenopathy, hepatosplenomegaly, cytopenias, and rash. Here we present a case of a 62-year-old male with progressive cervical adenopathy, fevers and weight loss presenting with extreme polyclonal plasmacytosis and high plasma EBV viral load. While the initial presentation appeared to mimic plasma cell leukemia or severe infection, lymph node biopsy and bone marrow biopsy confirmed a diagnosis of AITL. This case highlights the heterogeneity of …

Gm1 Ganglioside Modifies Α-Synuclein Toxicity And Is Neuroprotective In A Rat Α-Synuclein Model Of Parkinson's Disease., Jay S. Schneider, Radha Aras, Courtney K. Williams, James B. Koprich, Jonathan M. Brotchie, Vikrant Singh

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

While GM1 may interact with α-synuclein in vitro to inhibit aggregation, the ability of GM1 to protect against α-synuclein toxicity in vivo has not been investigated. We used targeted adeno-associated viral vector (AAV) overexpression of human mutant α-synuclein (A53T) in the rat substantia nigra (SN) to produce degeneration of SN dopamine neurons, loss of striatal dopamine levels, and behavioral impairment. Some animals received daily GM1 ganglioside administration for 6 weeks, beginning 24 hours after AAV-A53T administration or delayed start GM1 administration for 5 weeks beginning 3 weeks after AAV-A53T administration. Both types of GM1 administration protected against loss of SN …

The Top 10 Things To Know About Transfusion Medicine Before Intern Year: An Evidence-Based Course For Graduating Medical Students., Alexis R. Peedin, Irina Perjar, Marshall A. Mazepa, Marian A. Rollins-Raval, Yara A. Park, Jay S. Raval

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Background: Transfusion medicine (TM) knowledge varies widely among physician trainees. In addition, there have been few instances in which curricular changes have been meaningfully assessed for TM education in medical school.

Methods: We created and presented a novel lecture to improve TM knowledge for graduating medical students using eight objectives designed to reinforce critical information about blood management. Each objective was coded according to unique color schemes, fonts, and graphics to create visual associations while quickly and clearly presenting complex concepts. The validated BEST Collaborative exam was used to measure changes in student TM knowledge, while a survey was conducted …

Second-Harmonic Generation Microscopy Analysis Reveals Proteoglycan Decorin Is Necessary For Proper Collagen Organization In Prostate., Kirby R. Campbell, Rajeev Chaudhary, Monica Montano, Renato V. Iozzo, Wade A. Bushman, Paul J. Campagnola

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Collagen remodeling occurs in many prostate pathologies; however, the underlying structural architecture in both normal and diseased prostatic tissues is largely unexplored. Here, we use second-harmonic generation (SHG) microscopy to specifically probe the role of the proteoglycan decorin (Dcn) on collagen assembly in a wild type (wt) and Dcn null mouse (Dcn  -    /    -  ). Dcn is required for proper organization of collagen fibrils as it regulates size by forming an arch-like structure at the end of the fibril. We have utilized SHG metrics based on emission directionality (forward-backward ratio) and relative conversion efficiency, which are both related to …

Is Excision Of Radial Scars Identified On Cnb Necessary?, K. Nimtz, K. Hookim, Md, A. Sevrukov, Md, T. Tsangaris, Md, A. Willis, Md, A. Berger, Md, M. Lazar, Md

Phase 1

Introduction: Quantifying the risk of upgrade to malignancy with radial scars has been an ongoing challenge, as the published upgrade rate varies widely from 0-40%, making management strategy controversial. The lack of consensus on optimal management highlights the need for further analysis. We sought to identify our institutional upgrade rate of radial scar identified on core needle biopsy (CNB).

Methods: A retrospective review of pathology and radiology databases was performed to identify radial scars found on CNB. We excluded patients with malignancy associated with radial scar and those who did not undergo surgical excision. The upgrade rates to …

High Density Of Tumor-Associated Macrophage Staining Correlates With Poor Clinicopathologic Markers In Head And Neck Squamous Cell Carcinoma: A Meta-Analysis, Alexander Knops, Ba, Ayan Kumar, Bs, Brian Swendseid, Md, Ubaldo E. Martinez-Outshoorn, Md, Larry Harshyne, Phd, Nancy Philp, Phd, Ulrich Rodeck, Md, Phd, Christopher Snyder, Adam Luginbuhl, Md, David Cognetti, Md, Jennifer Johnson, Md, Joseph Curry, Md

Phase 1

Background: Head and neck squamous cell carcinoma (HNSCC) develops within a complex cellular microenvironment that promotes tumor growth, but also represents many potential therapeutic targets. Macrophage presence within that environment has been implicated in the growth, aggression, and persistence of HNSCC. Current literature reports variable degrees of association between tumor-associated macrophage (TAMs) density and clinicopathologic markers of disease.Inconsistent findings may result from grouping of TAM subtypes, which include both M1 (pro-inflammatory) and M2 (immunosuppressive). Our aim is to define the prognostic significance of the phenotypes of tumor-associated macrophages in HNSCC.

Methods: We conducted a meta-analysis of the existing publications investigating …

Cellular Milieu Imparts Distinct Pathological Α-Synuclein Strains In Α-Synucleinopathies, Ronald Gathagan, Chao Peng, Dustin Covell, Coraima Medellin, Anna Stieber, John L. Robinson, Bin Zhang, Rose M. Pitkin, Modupe F. Olufemi, Kelvin C. Luk, John Q. Trojanowski, Virginia M.Y. Lee

Phase 1

Introduction: In Lewy body diseases-including Parkinson's disease, without or with dementia, dementia with Lewy bodies, and Alzheimer's disease with Lewy body co-pathology -α-synuclein (α-Syn) aggregates in neurons as Lewy bodies and Lewy neurites. By contrast, in multiple system atrophy α-Syn accumulates mainly in oligodendrocytes as glial cytoplasmic inclusions (GCIs)

Objective: Our objective was to determine the conformational and biological profiles of a-Syn strains.

Methods: The following methods were used to collect and analyze data: Recombinant α-Syn purification and in vitro fibrillization. Preparation of sarkosyl-insoluble fractions from disease and control brains. Sandwich ELISA. Cell cultures. Stereotaxic injection of sarkosyl-insoluble fraction of …

Model-Based Virtual Patient Analysis Of Human Liver Regeneration Predicts Critical Perioperative Factors Controlling The Dynamic Mode Of Response To Resection., Babita Verma, Pushpavanam Subramaniam, Rajanikanth Vadigepalli

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

BACKGROUND: Liver has the unique ability to regenerate following injury, with a wide range of variability of the regenerative response across individuals. Existing computational models of the liver regeneration are largely tuned based on rodent data and hence it is not clear how well these models capture the dynamics of human liver regeneration. Recent availability of human liver volumetry time series data has enabled new opportunities to tune the computational models for human-relevant time scales, and to predict factors that can significantly alter the dynamics of liver regeneration following a resection.

METHODS: We utilized a mathematical model that integrates signaling …

Assessing The Utilization Of High-Resolution 2-Field Hla Typing In Solid Organ Transplantation., Yanping Huang, Anh Dinh, Steven Heron, Allison Gasiewski, Carolina Kneib, Hilary Mehler, Michael T. Mignogno, Ryan Morlen, Larissa Slavich, Ethan Kentzel, Edward C. Frackelton, Jamie L. Duke, Deborah Ferriola, Timothy Mosbruger, Olga A. Timofeeva, Steven S. Geier, Dimitri Monos

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

HLA typing in solid organ transplantation (SOT) is necessary for determining HLA-matching status between donor-recipient pairs and assessing patients' anti-HLA antibody profiles. Histocompatibility has traditionally been evaluated based on serologically defined HLA antigens. The evolution of HLA typing and antibody identification technologies, however, has revealed many limitations with using serologic equivalents for assessing compatibility in SOT. The significant improvements to HLA typing introduced by next-generation sequencing (NGS) require an assessment of the impact of this technology on SOT. We have assessed the role of high-resolution 2-field HLA typing (HR-2F) in SOT by retrospectively evaluating NGS-typed pre- and post-SOT cases. HR-2F …

Loss-Of-Function Mutations In Atp6ap1 And Atp6ap2 In Granular Cell Tumors., Fresia Pareja, Alissa H. Brandes, Thais Basili, Pier Selenica, Felipe C. Geyer, Dan Fan, Arnaud Da Cruz Paula, Rahul Kumar, David N. Brown, Rodrigo Gularte-Mérida, Barbara Alemar, Rui Bi, Raymond S. Lim, Ino De Bruijn, Sho Fujisawa, Rui Gardner, Elvin Feng, Anqi Li, Edaise M. Da Silva, John R. Lozada, Pedro Blecua, Leona Cohen-Gould, Achim A. Jungbluth, Emad A. Rakha, Ian O. Ellis, Maria I.A. Edelweiss, Juan P. Palazzo, Larry Norton, Travis Hollmann, Marcia Edelweiss, Brian P. Rubin, Britta Weigelt, Jorge S. Reis-Filho

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Granular cell tumors (GCTs) are rare tumors that can arise in multiple anatomical locations, and are characterized by abundant intracytoplasmic granules. The genetic drivers of GCTs are currently unknown. Here, we apply whole-exome sequencing and targeted sequencing analysis to reveal mutually exclusive, clonal, inactivating somatic mutations in the endosomal pH regulators ATP6AP1 or ATP6AP2 in 72% of GCTs. Silencing of these genes in vitro results in impaired vesicle acidification, redistribution of endosomal compartments, and accumulation of intracytoplasmic granules, recapitulating the cardinal phenotypic characteristics of GCTs and providing a novel genotypic-phenotypic correlation. In addition, depletion of ATP6AP1 or ATP6AP2 results in …

Mutant Ubqln2p497h In Motor Neurons Leads To Als-Like Phenotypes And Defective Autophagy In Rats, Tianhong Chen, Bo Huang, Xinglong Shi, Limo Gao, Cao Huang

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Mutations in ubiquilin2 (UBQLN2) have been linked to abnormal protein aggregation in amyotrophic lateral sclerosis (ALS). The mechanisms underlying UBQLN2-related neurodegenerative diseases remain unclear. Using a tetracycline-regulated gene expression system, the ALS-linked UBQLN2P497H mutant was selectively expressed in either the spinal motor neurons or astrocytes in rats. We found that selectively expressing mutant UBQLN2P497H in the spinal motor neurons caused several core features of ALS, including the progressive degeneration of motor neurons, the denervation atrophy of skeletal muscles, and the abnormal protein accumulation. Furthermore, mutant UBQLN2P497H accumulation was associated with an age-dependent decrease in several core autophagy-related proteins. ALS-like phenotypes …

Hypoxia-Mediated Mitochondria Apoptosis Inhibition Induces Temozolomide Treatment Resistance Through Mir-26a/Bad/Bax Axis., Xin Ge, Min-Hong Pan, Lin Wang, Wei Li, Chengfei Jiang, Jun He, Khaled Abouzid, Ling-Zhi Liu, Zhumei Shi, Bing-Hua Jiang

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Glioblastoma multiforme (GBM) is one of the most hypoxic tumors of the central nervous system. Although temozolomide (TMZ) is an effective clinical agent in the GBM therapy, the hypoxic microenvironment remains a major barrier in glioma chemotherapy resistance, and the underlying mechanisms are poorly understood. Here, we find hypoxia can induce the protective response to mitochondrion via HIF-1α-mediated miR-26a upregulation which is associated with TMZ resistance in vitro and in vivo. Further, we demonstrated that HIF-1α/miR-26a axis strengthened the acquisition of TMZ resistance through prevention of Bax and Bad in mitochondria dysfunction in GBM. In addition, miR-26a expression levels negatively …

Potential Role Of Csf Cytokine Profiles In Discriminating Infectious From Non-Infectious Cns Disorders., Danielle Fortuna, D. Craig Hooper, Amity L. Roberts, Larry A. Harshyne, Michelle Nagurney, Mark T. Curtis

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Current laboratory testing of cerebrospinal fluid (CSF) does not consistently discriminate between different central nervous system (CNS) disease states. Rapidly distinguishing CNS infections from other brain and spinal cord disorders that share a similar clinical presentation is critical. New approaches focusing on aspects of disease biology, such as immune response profiles that can have stimulus-specific attributes, may be helpful. We undertook this preliminary proof-of-concept study using multiplex ELISA to measure CSF cytokine levels in various CNS disorders (infections, autoimmune/demyelinating diseases, lymphomas, and gliomas) to determine the potential utility of cytokine patterns in differentiating CNS infections from other CNS diseases. Both …

Bacterial Contamination Of Platelets For Transfusion: Strategies For Prevention., Jerrold H. Levy, Matthew D. Neal, Jay H. Herman

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Platelet transfusions carry greater risks of infection, sepsis, and death than any other blood product, owing primarily to bacterial contamination. Many patients may be at particular risk, including critically ill patients in the intensive care unit. This narrative review provides an overview of the problem and an update on strategies for the prevention, detection, and reduction/inactivation of bacterial contaminants in platelets. Bacterial contamination and septic transfusion reactions are major sources of morbidity and mortality. Between 1:1000 and 1:2500 platelet units are bacterially contaminated. The skin bacterial microflora is a primary source of contamination, and enteric contaminants are rare but may …