Autopsy Specimens Research Articles

The Tumor-Suppressive Human Circular RNA CircITCH Sponges miR-330-5p to Ameliorate Doxorubicin-Induced Cardiotoxicity Through Upregulating SIRT6, Survivin, and SERCA2a.

Doxorubicin is one of the most potent antitumor agents available; however, its clinical use is restricted because it poses a risk of severe cardiotoxicity. Previous work has established that CircITCH (circular RNA ITCH [E3 ubiquitin-protein ligase]) is a broad-spectrum tumor-suppressive circular RNA and that its host gene, ITCH (E3 ubiquitin protein ligase), is involved in doxorubicin-induced cardiotoxicity (DOXIC). Whether CircITCH plays a role in DOXIC remains unknown. We aimed to dissect the role of CircITCH in DOXIC and further decipher its potential mechanisms. Circular RNA sequencing was performed to screen the potentially involved circRNAs in DOXI pathogenesis. Quantitative polymerase chain reaction and RNA in situ hybridization revealed that CircITCH was downregulated in doxorubicin-treated human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as well as in the autopsy specimens from cancer patients who suffered from doxorubicin-induced cardiomyopathy. Cell death/viability assays, detection of cardiomyocyte necrosis markers, microelectrode array, and cardiomyocyte functional assays revealed that CircITCH ameliorated doxorubicin-induced cardiomyocyte injury and dysfunction. Detection of cellular/mitochondrial oxidative stress and DNA damage markers verified that CircITCH alleviated cellular/mitochondrial oxidative stress and DNA damage induced by doxorubicin. RNA pull-down assays, Ago2 immunoprecipitation and double fluorescent in situ hybridization identified miR-330-5p as a direct target of CircITCH. Moreover, CircITCH was found to function by acting as an endogenous sponge that sequestered miR-330-5p. Bioinformatic analysis, luciferase reporter assays, and quantitative polymerase chain reaction showed that SIRT6 (sirtuin 6), BIRC5 (baculoviral IAP repeat containing 5, Survivin), and ATP2A2 (ATPase sarcoplasmic/endoplasmic reticulum Ca2+ transporting 2, SERCA2a [SR Ca2+-ATPase 2]) were direct targets of miR-330-5p and that they were regulated by the CircITCH/miR-330-5p axis in DOXIC. Further experiments demonstrated that CircITCH-mediated alleviation of DOXIC was dependent on the interactions between miR-330-5p and the 3'-UTRs of SIRT6, BIRC5, and ATP2A2 mRNA. Finally, AAV9 (adeno-associated virus serotype 9) vector-based overexpression of the well-conserved CircITCH partly prevented DOXIC in mice. CircITCH represents a novel therapeutic target for DOXIC because it acts as a natural sponge of miR-330-5p, thereby upregulating SIRT6, Survivin and SERCA2a to alleviate doxorubicin-induced cardiomyocyte injury and dysfunction.

The anatomic variations and surgical windows among optic chiasm/nerves and carotid arteries in the sellar region play a role in choosing the best surgical approaches: A Cadaveric study

ObjectiveUnderstanding the relationship between the carotid artery, optic nerve and the anterior clinoid process is the basis of surgical approaches performed in the subchiasmal region. The location of the optic chiasm, the length of the optic nerves, and the distance and angle between the optic nerves determine the route of surgical approaches. We have determined the types of optic chiasm to study the relationship between vascular and neural structures in this region. Materials and methodsThirty autopsy specimens were investigated at the Bursa Forensic Medicine Institute for optic chiasm types and the relationship between the neural and vascular anatomical structures of the sellar–parasellar and subchiasmal region was examined between June 2016 and November 2016. ResultsIn this study, 4 prefix types (13%), 6 postfix types (20%), and 20 central types (67%) of chiasm were defined. Furthermore, we measured this angle between two optic nerves, which indirectly shows the location of chiasm according to the diaphragma sellae, and then detected the mean value of this angle as 87.1±11.6°. The “limit” value to designate a chiasm as prefix was measured in the current study as ≥101.1°. The angle between optic nerves ranged from a mean value of 69.9±3.7° in 6 cases with postfix chiasm, to a mean value of 104.0±2.1° in 4 cases with prefix chiasm and a mean value of 88.8±6.7° in 20 cases with central chiasm. ConclusionIn this study, we showed that the relationship among optic chiasma types, optic nerves and bony and vascular structures around the sellar area was effective at determining the surgical approach to this region.

Tumor and tumor like lesions detected in autopsy specimens- A six year study done in a tertiary care hospital

Tumor and tumor like lesions detected in autopsy specimens- A six year study done in a tertiary care hospital

Targeting potential drivers of COVID-19: Neutrophil extracellular traps.

Coronavirus disease 2019 (COVID-19) is a novel, viral-induced respiratory disease that in ∼10-15% of patients progresses to acute respiratory distress syndrome (ARDS) triggered by a cytokine storm. In this Perspective, autopsy results and literature are presented supporting the hypothesis that a little known yet powerful function of neutrophils-the ability to form neutrophil extracellular traps (NETs)-may contribute to organ damage and mortality in COVID-19. We show lung infiltration of neutrophils in an autopsy specimen from a patient who succumbed to COVID-19. We discuss prior reports linking aberrant NET formation to pulmonary diseases, thrombosis, mucous secretions in the airways, and cytokine production. If our hypothesis is correct, targeting NETs directly and/or indirectly with existing drugs may reduce the clinical severity of COVID-19.

Intrathoracic Liver Heterotopia: An Incidental Finding at Autopsy

AbstractHeterotopia is the presence of a particular tissue at a nonphysiological site. The most common sites are gastric, pancreatic, and splenic heterotopias. While liver heterotopias are rare, intrathoracic heterotopias are much more limited. It is usually reported incidentally on imaging investigations or at autopsy. There is a high incidence of malignant transformation of heterotopic livers, and hence resection is always recommended. Here we report a case of heterotopic liver tissue in the right lung in a 42-year-old male, incidentally found at the gross examination of autopsy specimen of the lung.

MP26-19 TESTICULAR HISTOPATHOLOGY AFTER IMMUNOTHERAPY WITH IMMUNE CHECKPOINT INHIBITORS - A RETROSPECTIVE COHORT STUDY

You have accessJournal of UrologyInfertility: Epidemiology & Evaluation I (MP26)1 Apr 2020MP26-19 TESTICULAR HISTOPATHOLOGY AFTER IMMUNOTHERAPY WITH IMMUNE CHECKPOINT INHIBITORS - A RETROSPECTIVE COHORT STUDY Taylor P Kohn*, Jason Scovell, Iryna Samarska, Mahir Maruf, Eleonora Duregon, Jody Hooper, Shamlal Mangrav, Andres Matoso, and Amin S Herati Taylor P Kohn*Taylor P Kohn* More articles by this author , Jason ScovellJason Scovell More articles by this author , Iryna SamarskaIryna Samarska More articles by this author , Mahir MarufMahir Maruf More articles by this author , Eleonora DuregonEleonora Duregon More articles by this author , Jody HooperJody Hooper More articles by this author , Shamlal MangravShamlal Mangrav More articles by this author , Andres MatosoAndres Matoso More articles by this author , and Amin S HeratiAmin S Herati More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000865.019AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Immune checkpoint inhibitors (ICIs) are frequently used in the treatment of metastatic melanoma, clear cell renal cell carcinoma, and an increasing number of other cancers. Approximately 5% of these cancers will be diagnosed in adults of reproductive age. While chemotherapy is known to be gonadotoxic, no study has assessed the impact of ICI on spermatogenesis and the majority of preclinical data was performed on animals that were not yet sexually mature. The objective of this study is to assess the histologic impact of ICI on human spermatogenesis using autopsy specimens comparing testicular histology. METHODS: Men with a history of melanoma who had undergone immunotherapy for > 1 month with ipilimumab, nivolumab, and pembrolizumab between the years of 2013 and 2016 were included. A 1:1 proportioned control cohort was created and matched for age at biopsy. Men with a history of systemic chemotherapy or radiation were excluded. Tissue specimens were retrospectively examined from both living patients with testicular biopsies and deceased patients with testicular autopsies. For each testis specimen Johnsen score, seminiferous tubule density, Leydig cell density, and seminiferous tubule diameter were assessed. Medians were compared using Mann-Whitney U test using R 3.4.1. RESULTS: A total of seven men were included in the immunotherapy group with a median age at time of tissue biopsy of 58 years (range 23-66). Nine men were included in the control group with a median age at time of biopsy of 57 years (range 23-83). For patients undergoing autopsy, median time to autopsy was 7 hours in the ICI group and 6 hours in the control group. No difference was seen in Johnsen score when comparing ICI cohort and the control group, 8 (range: 2-10) and 9 (range: 2-10), respectively (p=0.55). No significant difference was seen between the ICI group and controls when assessing the number of seminiferous tubules per 200 um (11 vs 12), Leydig cells per 100 um (30 vs 26), or seminiferous tubular diameter (142 vs 171 um), respectively (p>0.05). CONCLUSIONS: In this retrospective cohort study assessing the impact of ICI, we found no significant difference in testicular histology between those receiving ICI and those who had never received ICI. This null finding suggests that ICI agents may not be spermatotoxic, unlike chemotherapy counterparts, and may be a valuable treatment when fertility preservation is desired. Large multicenter registries are needed to further track potential side-effects associated with ICI. Source of Funding: None © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e408-e408 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Taylor P Kohn* More articles by this author Jason Scovell More articles by this author Iryna Samarska More articles by this author Mahir Maruf More articles by this author Eleonora Duregon More articles by this author Jody Hooper More articles by this author Shamlal Mangrav More articles by this author Andres Matoso More articles by this author Amin S Herati More articles by this author Expand All Advertisement PDF downloadLoading ...

Morbid Anatomy of Patients with Mycobacterial Infections, a Cross Sectional Autopsy Study at a Tertiary Hospital in a High Disease Burden Setting

Background: Tuberculosis is a major cause of morbidity and mortality globally. Infections caused by different mycobacteria can be clinically indistinguishable but the differences in their pathological features and mortality are unclear. Methods: To evaluate the contribution of different mycobacterial pathogens to lethal mycobacterial disease and assess subsequent pathological features, we performed autopsies of 49 patients with suspected TB. Autopsy specimens were examined histologically and cultured for mycobacteria. We identified Mycobacterium tuberculosis ( Mtb ) isolates and non-tuberculous mycobacteria (NTM), and further genotyped the Mtb isolates. Results: Mtb isolates were found in 37 patients and NTM in 12 patients. All patients had signs of caseous pneumonia and 42 patients (86%) had disease involving more than half of the lungs. Two-thirds of the patients with TB or NTM had extrapulmonary engagement in addition to pulmonary pathology. Gross pathology and histopathology were similar in TB patients and patients with NTM, except that patients with NTM had a significantly reduced tendency to cause pleural effusions. Of the Mtb isolates 55% were of the Uganda genotype, which is the predominant Mtb genotype in Uganda. This genotype was significantly more frequent in the younger patients. Conclusion: In this autopsy study of patients with presumptive TB a majority of culture positive cases were caused by Mtb. In 25% of patients NTM were identified, and histopathology was similar in TB and NTM patients, although NTM patients had significantly less pleural effusions. These results suggest that NTM contribute to mortality, and that their identification is important because of the special clinical and therapeutic implications associated with NTM infections. Funding Statement: The study was supported by funds from the Swedish International Development Cooperation Agency through Makerere University-Karolinska Institutet Research collaboration, and the Swedish Heart-Lung Foundation. Declaration of Interest: The authors declare that they have no competing interests. Ethical Approval Statement: The study was approved by the Internal Review Board (IRB) of the Makerere University School of Medicine. Final approval was granted by the Uganda National Council of Science and Technology. (HS 1364).

Tumor Location and Pathological Features of Latent and Incidental Prostate Cancer in Contemporary Japanese Men.

The incidence of prostate cancer is increasing in Asian countries. Studies using prostatectomy specimens have reported a racial difference in tumor location within the prostate, with a greater incidence of transition zone cancer in Asian men compared with Caucasian men. However, there may be potential biases in studies based on surgical specimens. We describe the pathological features of subclinical prostate cancer, such as latent cancer and incidental cancer, to elucidate tumor location of contemporary Japanese patients. We also compare the prevalence of latent and incidental prostate cancer to determine whether the incidence of prostate cancer is higher in patients with bladder cancer. Overall 182 men autopsied and 148 who underwent cystoprostatectomy for bladder cancer were included in the study. Each prostate gland was fixed and sliced in step sections. Histological evaluation was performed by a single genitourinary pathologist. The index tumor location was categorized into transition zone or peripheral zone. Prostate cancer was found in 39.0% of the autopsy specimens and 31.6% of the cystoprostatectomy specimens. The prevalence and pathological characteristics were not significantly different between latent and incidental cancer. The prevalence of transition zone cancer was 39.0% (46 of 118). In elderly men peripheral zone cancer was more frequently diagnosed than transition zone cancer (p=0.049). The pathological characteristics of transition and peripheral zone cancers were similar except for the pT stage. Transition zone cancer was prevalent in contemporary Japanese men. The incidence of prostate cancer in men with bladder cancer might not be higher than that in healthy men.

Brain tissue gadolinium retention in pediatric patients after contrast-enhanced magnetic resonance exams: pathological confirmation.

Retained gadolinium from gadolinium-based contrast agents (GBCAs) used in MR exams has been inferred based on signal changes on serial brain MRI and subsequently demonstrated pathologically in adults. Retention has been similarly inferred in children but pathological demonstration in pediatric patients is limited. The long-term effects of retained gadolinium are unknown but are potentially of greater concern in children given their increased vulnerability from continuing development and their expected longer period of exposure. Several factors can influence gadolinium retention. In adults as well as in children, greater accumulation has been demonstrated based on MR signal changes with linear compared with macrocyclic gadolinium chelates, attributed to lower chelate affinity with linear agents. Effects of age at exposure on retention are unknown, while differences in GBCA washout rates are still under investigation and might affect gadolinium retention relative to time of GBCA administration. The purpose of this study was to confirm whether gadolinium brain deposits are present in pediatric patients who received GBCAs and to quantify the amounts present. Brain autopsy specimens from 10 pediatric patients between 1year and 13years of age who underwent at least one contrast-enhanced MR exam were analyzed for elemental gadolinium using inductively coupled plasma mass spectrometry. Brain samples included white matter, basal ganglia (putamen, globus pallidus), thalamus, dentate nucleus and tumor tissue as available. Type and dose of contrast agent, number and timing of contrast-enhanced MR exams and renal function (estimated glomerular filtration rate [eGFR]) were documented for each child. Patient exposures ranged from 1 dose to 20 doses of GBCAs including both macrocyclic and linear ionic agents. Gadolinium was found to be present in brain tissue in all children and was generally highest in the globus pallidus. Those who received only macrocyclic agents showed lower levels of gadolinium retention. This study demonstrates pathological confirmation of gadolinium retention in brain tissue of a series of pediatric patients exposed to GBCAs including not only linear ionic agents but also macrocyclic agents with both nonionic and ionic compounds. The distribution and deposition levels in this small pediatric population are comparable with the findings in adults. While the clinical significance of these deposits remains unknown, at this point it would be prudent to exert caution and avoid unnecessary use of GBCAs in pediatric patients.

Francisella tularensis infection in dogs: 88 cases (2014-2016).

To characterize the epidemiology, clinical signs, and treatment of dogs with Francisella tularensis infection in New Mexico. 87 dogs in which 88 cases of tularemia (1 dog had 2 distinct cases) were confirmed by the New Mexico Department of Health Scientific Laboratory Division from 2014 through 2016 and for which medical records were available. Dogs were confirmed to have tularemia if they had a 4-fold or greater increase in anti-F tularensis antibody titer between acute and convalescent serum samples or F tularensis had been isolated from a clinical or necropsy specimen. Epidemiological, clinical, and treatment information were collected from the dogs' medical records and summarized. All 88 cases of tularemia were confirmed by paired serologic titers; the first (acute) serologic test result was negative for 84 (95%) cases. The most common reported exposure to F tularensis was wild rodent or rabbit contact (53/88 [60%]). Dogs had a median number of 3 clinical signs at initial evaluation; lethargy (81/88 [92%]), pyrexia (80/88 [91%]), anorexia (67/88 [76%]), and lymphadenopathy (18/88 [20%]) were most common. For 32 (36%) cases, the dog was hospitalized; all hospitalized dogs survived. Dogs with F tularensis infection often had nonspecific clinical signs and developed moderate to severe illness, sometimes requiring hospitalization. Veterinarians examining dogs from tularemia-enzootic areas should be aware of the epidemiology and clinical signs of tularemia, inquire about potential exposures, and discuss prevention methods with owners, including reducing exposure to reservoir hosts and promptly seeking care for ill animals.

Neonatal Autopsies with Heart Disease: A Challenge for Pathologist

Congenital Heart Diseases (CHD) carry significant morbidity and mortality in paediatric patients. Transposition of Great Arteries (TGA) is a common cyanotic CHD. However, onset of Pulmonary Arterial Hypertension (PAH) is potentially severe and rare complication reported in 1-3% of newborns. Hypertrophic Cardiomyopathy (HOCM) is a primary disease of cardiac muscle usually recognised in adulthood. Neonatal HOCM without other congenital anomalies, no family history of HOCM, no history of exposure to corticosteroids or any inborn errors of metabolism is rarely recognised. Authors hereby report two cases of neonatal autopsy showing TGA with PAH (Grade 2) and another with HOCM without any primary cause received in our hospital. With this case report authors wish to stress on the importance of cardiac examination (heart and the associated vessels) in autopsy specimen which in turn requires training of pathologists in cardiac pathology along with routine fetal anomaly scanning in arriving successively at the final diagnosis and detecting the cause of death which helps in management of future pregnancies by the clinicians.

Risques professionnels chez les pathologistes : résultats d’une enquête française

The profession of pathologist exposes to various risks, notably infectious, physical and chemical. The objective of this study was to make an inventory of these occupational risks to which pathologists are subjected and to evaluate the pathologies that they presented. A particular attention was given to microscopic and screen work as they can induce musculoskeletal or ophthalmic disorders, and stress-related psychological disorders. An anonymous online questionnaire containing 54 questions had been sent by mail to pathologists via the French Society of Pathology and the Syndicate of French Pathologists. Five hundred and twelve pathologists responded to the survey. Thirty-eight percent reported musculoskeletal disorders in the last 6 months. Visual disturbances concerned 73.4% of respondents. In the last 12 months, 33.3% of pathologists had been injured or had had mucosal projections during macroscopic or autopsy specimens. The frequency of infectious diseases was low (6.2%) as well as that of cancers (3.9%). Psychological disorders such as depression or burnout were reported by 16.7% of respondents. Pathologists seemed happy at work and had a good overall lifestyle. Few doctors had medical follow-up and few had benefited from ergonomic advice and training on the risks of chemicals. The results of this study showed the interest of a medical surveillance adapted to the pathologies presented by the pathologists. Educational and preventive measures should be introduced early in the career, focusing on ergonomics and learning about chemical and biological hazards.

Detection and Quantification of Screw-Home Movement Using Nine-Axis Inertial Sensors

Purpose: Although previous studies on the screw-home movement (SHM) for autopsy specimen and walking of living persons conduc­t ed, the possibility of acquiring SHM based on inertial measurement units received little attention. This study aimed to investigate the possibility of measuring SHM for the non-weighted bearing using a micro-electro-mechanical system-based wearable motion capture system (MEMSS). Methods: MEMSS and camera-based motion analysis systems were used to obtain kinematic data of the knee joint. The knee joint moved from the flexion position to a fully extended position and then back to the start point. The coefficient of multiple correlation and the difference in the range of motion were used to assess the waveform similarity in the movement measured by two measurement sys­ tems. Results: The waveform similarity in the sagittal plane was excellent and the in the transverse plane was good. Significant differences were found in the sagittal plane between the two systems (p 0.05). Conclusion: The SHM during the passive motion without muscle contraction in the non-weighted bearing appeared in the entire range. We thought that the MEMSS could be easily applied to the acquisition of biomechanical data on the knee related to physical therapy.

Cardiomyocyte-Specific JunD Overexpression Increases Infarct Size following Ischemia/Reperfusion Cardiac Injury by Downregulating Sirt3.

Ischemia/reperfusion (I/R) injury in acute myocardial infarction activates several deleterious molecular mechanisms. The transcription factor JunD regulates pathways involved in oxidative stress as well as in cellular proliferation, differentiation, and death. The present study investigated the potential role of JunD as a modulator of myocardial injury pathways in a mouse model of cardiac I/R injury. Infarct size, systemic and local inflammation, and production of reactive oxygen species, as well as cytosolic and mitochondrial apoptotic pathways were investigated in adult males after myocardial I/R. In wild-type (WT) mice, 30 minutes after ischemia and up to 24 hours following reperfusion, cardiac JunD messenger ribonucleic acid expression was reduced while JunB increased. Cardiac-specific JunD overexpressing mice (JunDTg/0 ) displayed larger infarcts compared with WT. However, postischemic inflammatory or oxidative responses did not differ. JunD overexpression reduced Sirt3 transcription by binding to its promoter, thus leading to mitochondrial dysfunction, myocardial cell death, and increased infarct size. On the other hand, JunD silencing reduced, while Sirt3 silencing increased infarct size. In human myocardial autopsy specimens, JunD-positive areas within the infarcted left ventricle staining corresponded to undetectable Sirt3 areas in consecutive sections of the same heart. Cardiac-specific JunD overexpression increases myocardial infarct size following I/R. These effects are mediated via Sirt3 transcriptional repression, mitochondrial swelling, and increased apoptosis, suggesting that JunD is a key regulator of myocardial I/R injury. The present data set the stage for further investigation of the potential role of Sirt3 activation as a novel target for the treatment of acute myocardial infarction.

Clinical, pathological and therapeutic findings on lingual rhabdomyosarcoma in a dog - case report

ABSTRACT The biological behaviour of the tumours vary according to the species in which they occur, its location within the cavity, clinical stage and histopathological nature. Lingual neoplasms are generally uncommon in companion animals. Rhabdomyosarcomas are malignant, solid, aggressive formations with high metastatic potential. The clinical signs are variable and a definitive diagnosis can only be reached through histopathological analysis of biopsy and necropsy specimen. In some cases, immunohistochemical study may be needed to confirm the diagnosis. This paper aims to highlight important points about this uncommon condition in dogs, using a case report of lingual rhabdomyosarcoma, which showed no evidence of metastasis after diagnosis, nor of local recurrence after surgical excision with a wide safety margin. It was concluded that early diagnosis, the correct interpretation of the complementary tests and the appropriate therapeutic approach contributed to improving the quality of life and survival of the patient in question.

The anatomic variations and surgical windows among optic chiasm/nerves and carotid arteries in the sellar region play a role in choosing the best surgical approaches: A Cadaveric study.

Understanding the relationship between the carotid artery, optic nerve and the anterior clinoid process is the basis of surgical approaches performed in the subchiasmal region. The location of the optic chiasm, the length of the optic nerves, and the distance and angle between the optic nerves determine the route of surgical approaches. We have determined the types of optic chiasm to study the relationship between vascular and neural structures in this region. Thirty autopsy specimens were investigated at the Bursa Forensic Medicine Institute for optic chiasm types and the relationship between the neural and vascular anatomical structures of the sellar-parasellar and subchiasmal region was examined between June 2016 and November 2016. In this study, 4 prefix types (13%), 6 postfix types (20%), and 20 central types (67%) of chiasm were defined. Furthermore, we measured this angle between two optic nerves, which indirectly shows the location of chiasm according to the diaphragma sellae, and then detected the mean value of this angle as 87.1±11.6°. The "limit" value to designate a chiasm as prefix was measured in the current study as ≥101.1°. The angle between optic nerves ranged from a mean value of 69.9±3.7° in 6 cases with postfix chiasm, to a mean value of 104.0±2.1° in 4 cases with prefix chiasm and a mean value of 88.8±6.7° in 20 cases with central chiasm. In this study, we showed that the relationship among optic chiasma types, optic nerves and bony and vascular structures around the sellar area was effective at determining the surgical approach to this region.

Potential Genetic and Immunologic Mechanisms of Therapeutic Resistance and Disease Progression in CNS Lymphoma Elucidated Via Whole Brain Autopsy Studies

While the genomic landscape of CNS lymphomas at diagnosis has largely been defined, there remains a significant gap in our understanding of the molecular pathways that mediate therapeutic resistance and disease progression. CNS lymphomas generally exhibit multifocal dissemination as an end-stage manifestation. To elucidate key genetic and pathophysiologic pathways of CNS lymphoma that mediate resistance, we have conducted what may be the first study to apply next generation sequencing to compare the genetic features of PCNSL and SCNSL at the time of diagnosis with multifocal lesions isolated from relapsed, treatment-refractory disease from matched patient specimens at final lymphoma progression, isolated from whole brain autopsy specimens. Whole-exome sequencing from diagnostic specimens (4 brain and 1 retinal biopsy) were compared with matched whole brain autopsy specimens in 5 patients: 4 with PCNSL and 1 with SCNSL. A median of 6 distinct anatomical CNS lymphoma-containing brain regions were analyzed/whole brain autopsy specimen. Median age was 57 (range 51-81 yrs). Two patients died after primary refractory disease to methotrexate-based induction and three died after median of four progressions; each of these later patients progressed after lenalidomide, and one also received pomalidomide followed by ibrutinib before final disease progression. Four cases were from UCSF and 1 from Stanford. Tumor tissue and uninvolved normal brain was microdissected from formalin-fixed, paraffin-embedded blocks of initial biopsy prior to therapy and multiple distinct tumor regions at time of autopsy. Multiplex library preparation was performed from extracted genomic DNA and hybridization-capture of pooled libraries performed using NimbleGen SeqCap EZ Human Exome Probes Bait Library. Captured libraries were sequenced as paired-end 100 base pair reads and sequence reads mapped to the reference human genome build GRCh37 (hg19) using Sentieon DNAseq. Somatic variant detection including single nucleotide variants and insertions/deletions was performed. Single nucleotide variants and insertions/deletions were verified using Integrated Genome Viewer. Copy number analysis was performed using CNVkit and visualized using Nexus. High-confidence somatic nonsynonymous mutations were identified and pathway analysis applied using gene sets for canonical pathways and Gene Ontology (GO) terms downloaded from Molecular Signatures Database) at http://www.broad.mit.edu/gsea/msigdb. Lymphoid-specific signatures were curated by Staudt lab (http://lymphochip.nih.gov/signaturedb/) or previous publications. Fisher's Exact test was used to calculate enrichment p values for each of those gene sets and the BH method used for False Discovery Rate (FDR) control. A mean of 214 lymphoma-specific mutations per specimen was identified (range 56-1,764). Notably, of the 283 distinct molecular pathways analyzed, only 1 pathway was significantly enriched for mutations in tumors from each of the 5 CNS lymphoma whole brain autopsy specimens compared to pre-treatment diagnostic biopsies: the Ikaros 1(IKZF1) in centroblast pathway (p<0.05). The 2 cases of CNS lymphoma that had been exposed to an IMiD most proximate to brain autopsy (< 6 weeks) exhibited the strongest enrichment for mutations in IKZF1 pathway (p<0.005). Genes recurrently mutated in the IKZF1 pathway in relapsed CNS lymphomas included CD79A/B, IGLL5, BTG2, and TMEM30A. Mutations in the BCL6 and BACH2 transcriptional repressor pathway were significantly enriched in CNS lymphomas at autopsy compared to diagnostic specimens in 3 out of 5 cases. We also detected marked regional heterogeneity in DNA copy number aberrations in lymphoma from whole brain autopsy specimens. High level focal amplification of 9p24.1 targeting PD-L1 was enriched in only one case: a highly refractory PCNSL that had been exposed to pomalidomide followed by ibrutinib within two months of autopsy; amplification of PD-L1 was not detected in the diagnostic specimen. While multiple genetic aberrations are associated with disease progression in CNS lymphoma, our study suggests that mutational activation of the IKZF1 pathway may be a unique canonical mechanism in the evolution of resistance to multiple interventions, including first-line methotrexate. This observation may have significant therapeutic implications. Supported by NCI and Leukemia Lymphoma Society. Disclosures Rubenstein: Genentech: Research Funding; Celgene: Research Funding; Bristol Myers Squibb: Research Funding.

Zika virus infection: A correlation between prenatal ultrasonographic and postmortem neuropathologic changes

This study presents a correlation between prenatal ultrasonographic images and neuropathologic findings of postmortem tissue samples from five confirmed cases of perinatal Zika virus (ZIKV) infection belonging to the cohort of the ZEN Initiative in Bucaramanga, Colombia. Deaths occurred between June 2016 and March 2017. Mothers consulted with ZIKV infection clinical manifestations or fetal central nervous system (CNS) abnormalities or both. A detailed ultrasound scan and neurosonographic protocol was performed by maternal fetal specialists. Perinatal autopsies were performed following the Colombian National Health Institute's ZIKV protocol. The autopsies were from two fetal deaths, and three early neonatal deaths. Gestational age was between 262/7 and 382/7 weeks. Two cases were classified as mild microcephaly. Few findings by ultrasound and pathology were found in case 1 because it was a late infection; the other cases presented findings corresponding to congenital Zika syndrome: craniofacial malformations, cerebellar hypoplasia, anomalies of the corpus callosum and ventriculomegaly, all confirmed in autopsy specimens. By ultrasonography, hyperechogenicities were seen in several brain structures, which correspond to cortical and periventricular calcifications, subependymal glial reactivity and perivascular rings. The ultrasound and pathological findings show a wide spectrum of CNS anomalies that confirm the neurotropic effect of the ZIKV, recognizing the neuroimaging findings of this disease (unilateral ventriculomegaly, alterations in the corpus callosum and cerebellum, and calcifications) are highly suggestive of ZIKV infection.

GENE-53. IDENTIFICATION OF EPITHELIAL MEMBRANE PROTEIN 2 (EMP2) AS A MOLECULAR MARKER AND THERAPEUTIC TARGET IN MENINGIOMA

Abstract BACKGROUND Intracranial meningiomas have been associated with a heterogenous set of genetic and molecular alterations. Despite these genetic associations, targeted therapies have been unable to increase survival in recurrent or residual meningiomas. There is a need for identification of therapeutic targets in meningioma. MATERIALS & METHODS Normal brain tissue from routine autopsy and tumor specimens from patients undergoing primary surgical resection of meningioma were analyzed. Pathologic diagnosis and grade were identified. Samples were stained with anti-EMP2 antisera and visualized with DAB (3,3’-Diaminobenzidine). Western blot analysis was carried out RESULTS In 17 patients with meningioma, there was statistically significant EMP2 staining in fresh brain tumor samples. EMP2 expression was confirmed and quantified with western blot which demonstrated significant positive expressivity. There were 12 grade I, 4 grade II, and 1 grade III meningiomas in our dataset and a significant trend towards increased EMP2 expression with higher grade. EMP2 expression was not identified in normal brain tissue samples from different locations including gray matter, white matter, and meninges in 3 control patients. CONCLUSIONS EMP2 is a protein associated with cell migration, invasion, and angiogenesis not identified in normal brain tissue but found in meningioma. EMP2 expression may be associated with increased tumor invasion and is a potential therapeutic target for residual or recurrent meningioma.

Histological evidence for the cardiac safety of high-dose pegylated liposomal doxorubicin in a patient with HIV-associated Kaposi sarcoma: a case report and literature review

BackgroundPegylated liposomal doxorubicin plays an important role in the treatment of patients with severe refractory human immunodeficiency virus (HIV)-associated Kaposi sarcoma (KS). High cumulative doses of conventional doxorubicin exceeding 500 mg/m2 are known to cause cardiac toxicity. However, the safe cumulative dose of pegylated liposomal doxorubicin is unclear.Case presentationA 40-year-old Japanese man with HIV infection presented with pain, edema, and multiple skin nodules on both legs which worsened over several months. He was diagnosed with HIV-associated KS. He received long-term pegylated liposomal doxorubicin combined with antiretroviral therapy for advanced, progressive KS. The cumulative dose of pegylated liposomal doxorubicin reached 980 mg/m2. The patient’s left ventricular ejection fraction remained unchanged from baseline during treatment. After he died as a result of cachexia and wasting, caused by recurrent sepsis and advanced KS, an autopsy specimen of his heart revealed little or no evidence of histological cardiac damage. We also conducted a literature review focusing on histological changes of the myocardium in patients treated with a cumulative dose of pegylated liposomal doxorubicin exceeding 500 mg/m2.ConclusionsThis case report and literature review suggest that high (> 500 mg/m2) cumulative doses of pegylated liposomal doxorubicin may be used without significant histological/clinical cardiac toxicity in patients with HIV-associated KS.