Non-pathological Tissue Research Articles

Hsf1 and Hsf2 in normal, healthy human tissues: Immunohistochemistry provokes new questions.

The heat shock transcription factors heat shock transcription factor 1 and Hsf2 have been studied for many years, mainly in the context of stress response and in malignant cells. Their physiological function in nonmalignant human cells under nonstress conditions is still largely unknown. To approach this important issue, Joutsen et al. present immunohistochemical staining data on Hsf1 and Hsf2 in 80 nonpathological human tissue samples. The wealth of these data elicits many interesting questions that will spur many future research projects.

Comparative quantitation of liver-type fatty acid-binding protein localizations in liver injury and non-pathological liver tissue in dogs

Background and Aim: Liver injury results in the production of free radicals that can lead to hepatocytic degeneration, cirrhosis, and hepatocellular carcinoma (HCC). Liver-fatty acid-binding protein (L-FABP) is highly expressed in hepatocytes and is a key regulator of hepatic lipid metabolism and antioxidant characteristics. Interestingly, the increase in L-FABP expression could be used as a novel marker of liver injury. Therefore, this study aimed to use immunohistochemical techniques to investigate the expression of L-FABP in dogs with liver injury compared with dogs with non-pathological liver. Materials and Methods: Liver tissue samples were collected from dog biopsy specimens at the Veterinary Diagnostic Laboratory at the Faculty of Veterinary Medicine, Chiang Mai University. The tissues were prepared for immunohistochemistry and the expression and localization of L-FABP were investigated using one-way analysis of variance. Results: Immunohistochemical analysis showed that L-FABP was strongly expressed in the hepatocytes of dogs with lipidosis and HCC when compared with that in normal liver. Semi-quantitative immunohistochemistry evaluation showed the percentage of protein expression of L-FABP 0.023 ± 0.027 in the non-pathological liver. The percentage of L-FABP protein expression in lipidosis and HCC was found to be 8.517 ± 1.059 and 17.371 ± 4.026, respectively. Conclusion: L-FABP expression in dogs with liver injuries was significantly higher than that in dogs with non-pathological liver injury (p = 0.05). These results suggest that L-FABP has the potential as a novel marker for specific diagnosis and prognosis of dogs with liver injury. Keywords: canine, hepatocyte, immunohistochemistry, lipidosis, liver injury, liver-fatty acid-binding protein, pathology.

Identification of ferroptosis-related genes in acute kidney injury by bioinformatic analysis

ABSTRACT Background: Acute kidney injury (AKI) is a rapid decline in renal function characterized by a decrease in glomerular filtration rate in a short period. Ferroptosis affects the development of various kidney diseases. Therefore, searching for genes related to ferroptosis is crucial for understanding the occurrence and development of AKI. Methods: We obtained data from 18 samples (8 with acute kidney injury and 10 non-pathological tissue) in GSE53769 and 48 samples (39 with acute kidney injury and 9 non-pathological tissue) in GSE139061. We obtained differentially expressed genes (DEGs) of AKI/Control samples from both two datasets and intersected them with known ferroptosis-related genes (FRGs) to obtain ferroptosis-related DEGs (FRDEGs). GO annotation, KEGG pathway analysis, and GSEA analysis were conducted on the FRDEGs to understand their enriched biological functions and pathways. Next, we constructed the protein-protein interaction (PPI) network. Results: A total of 312 genes were obtained, which were abnormally expressed in both two datasets. After intersecting with known FRGs, 14 FRDEGs were obtained, namely ACSF2, ADIPOR1, ARF6, ATF3, ATF6, DPEP1, FH, GLRX5, MIOX, NAP1L1, NDRG1, PPARA, SPHK1, YY1AP1. The results of the immune infiltration analysis showed that multiple gene expressions among 14 genes are correlated with immune cell infiltration. Conclusion: Fourteen ferroptosis genes (ACSF2, ADIPOR1, ARF6, ATF3, ATF6, DPEP1, FH, GLRX5, MIOX, NAP1L1, NDRG1, PPARA, SPHK1, YY1AP1) are involved in the occurrence and development of AKI, among which NDRG1 might be the core functional gene, and PPARA is expected to become the most effective therapeutic target gene.

Non-invasive assessment of normal and impaired iron homeostasis in the brain

Strict iron regulation is essential for normal brain function. The iron homeostasis, determined by the milieu of available iron compounds, is impaired in aging, neurodegenerative diseases and cancer. However, non-invasive assessment of different molecular iron environments implicating brain tissue’s iron homeostasis remains a challenge. We present a magnetic resonance imaging (MRI) technology sensitive to the iron homeostasis of the living brain (the r1-r2* relaxivity). In vitro, our MRI approach reveals the distinct paramagnetic properties of ferritin, transferrin and ferrous iron ions. In the in vivo human brain, we validate our approach against ex vivo iron compounds quantification and gene expression. Our approach varies with the iron mobilization capacity across brain regions and in aging. It reveals brain tumors’ iron homeostasis, and enhances the distinction between tumor tissue and non-pathological tissue without contrast agents. Therefore, our approach may allow for non-invasive research and diagnosis of iron homeostasis in living human brains.

Temporal stability of intracranial electroencephalographic abnormality maps for localizing epileptogenic tissue.

Identifying abnormalities on interictal intracranial electroencephalogram (iEEG), by comparing patient data to a normative map, has shown promise for the localization of epileptogenic tissue and prediction of outcome. The approach typically uses short interictal segments of approximately 1 min. However, the temporal stability of findings has not been established. Here, we generated a normative map of iEEG in nonpathological brain tissue from 249 patients. We computed regional band power abnormalities in a separate cohort of 39 patients for the duration of their monitoring period (.92-8.62 days of iEEG data, mean = 4.58 days per patient, >4800 hours recording). To assess the localizing value of band power abnormality, we computed -a measure of how different the surgically resected and spared tissue was in terms of band power abnormalities-over time. In each patient, the value was relatively consistent over time. The median of the entire recording period separated seizure-free (International League Against Epilepsy [ILAE] = 1) and not-seizure-free (ILAE 1) patients well (area under the curve [AUC] = .69). This effect was similar interictally (AUC = .69) and peri-ictally (AUC = .71). Our results suggest that band power abnormality D_RS, as a predictor of outcomes from epilepsy surgery, is a relatively robust metric over time. These findings add further support for abnormality mapping of neurophysiology data during presurgical evaluation.

Spectral Similarity Measures for In Vivo Human Tissue Discrimination Based on Hyperspectral Imaging.

Similarity measures are widely used as an approved method for spectral discrimination or identification with their applications in different areas of scientific research. Even though a range of works have been presented, only a few showed slightly promising results for human tissue, and these were mostly focused on pathological and non-pathological tissue classification. In this work, several spectral similarity measures on hyperspectral (HS) images of in vivo human tissue were evaluated for tissue discrimination purposes. Moreover, we introduced two new hybrid spectral measures, called SID-JM-TAN(SAM) and SID-JM-TAN(SCA). We analyzed spectral signatures obtained from 13 different human tissue types and two different materials (gauze, instruments), collected from HS images of 100 patients during surgeries. The quantitative results showed the reliable performance of the different similarity measures and the proposed hybrid measures for tissue discrimination purposes. The latter produced higher discrimination values, up to 6.7 times more than the classical spectral similarity measures. Moreover, an application of the similarity measures was presented to support the annotations of the HS images. We showed that the automatic checking of tissue-annotated thyroid and colon tissues was successful in 73% and 60% of the total spectra, respectively. The hybrid measures showed the highest performance. Furthermore, the automatic labeling of wrongly annotated tissues was similar for all measures, with an accuracy of up to 90%. In future work, the proposed spectral similarity measures will be integrated with tools to support physicians in annotations and tissue labeling of HS images.

ANGI-10. IMPLICATIONS OF EPITHELIAL MEMBRANE PROTEIN-2 (EMP2) IN THE PATHOGENESIS OF MENINGIOMAS

Abstract INTRODUCTION VEGF-A is an important mediator of angiogenesis in meningiomas. However, targeting this protein with bevacizumab has offered only a modest improvement in survival, suggesting that there are other compensatory mechanisms involved, enabling vascularization despite ongoing treatment. Epithelial membrane protein 2 (EMP2), a cell surface protein, promotes angiogenic processes in gynecological cancers and brain tumors including gliomas and glioblastoma. However, little is known about its etiology and mechanism of action within meningiomas. OBJECTIVE We aimed to evaluate EMP2 expression in non-pathologic brain tissue, tumor specimens, and in meningioma cell lines. We also aimed to understand its mechanism within cells with regard to proliferation and invasion. METHODS Non-pathologic brain tissues from routine autopsies and tumor specimens from surgical resection were analyzed. Additionally, four meningioma cell lines were obtained (CH157, BEN-MEN1, SF1335 and IOMM-LEE) and changes in cellular proliferation and invasion were measured. Furthermore, EMP2 expression was modulated using lentiviral infection. A p-value < 0.05 was considered statistically significant. RESULTS EMP2 mRNA and protein expression were significantly higher in the meningioma specimens compared to non-pathologic meninges. Its expression also positively correlated with an angiomatous pattern and VEGF-A mRNA expression. In vitro, high EMP2 protein expression was noted in all cell lines, and quantification demonstrated highest expression in the malignant cell line, the IOMM-LEE. Viral vectors were subsequently utilized to create EMP2 overexpressing and knockdown IOMM-LEE models. Western analysis demonstrated a correlation between EMP2 overexpression and HIF1α, a key mediator of tumor growth and angiogenesis. CONCLUSION Our data from primary tumor samples and in vitro models implicate EMP2 in meningioma pathogenesis. This warrants further investigation by evaluating the effects of anti-EMP2 therapy on tumor growth and invasiveness in both in vitro andin vivo models.

Mapping the myelin bilayer with short-T2 MRI: Methods validation and reference data for healthy human brain.

To explore the properties of short-T2 signals in human brain, investigate the impact of various experimental procedures on these properties and evaluate the performance of three-component analysis. Eight samples of non-pathological human brain tissue were subjected to different combinations of experimental procedures including D2 O exchange and frozen storage. Short-T2 imaging techniques were employed to acquire multi-TE (33-2067 μs) data, to which a three-component complex model was fitted in two steps to recover the properties of the underlying signal components and produce amplitude maps of each component. For validation of the component amplitude maps, the samples underwent immunohistochemical myelin staining. The signal component representing the myelin bilayer exhibited super-exponential decay with T2,min of 5.48 μs and a chemical shift of 1.07ppm, and its amplitude could be successfully mapped in both white and gray matter in all samples. These myelin maps corresponded well to myelin-stained tissue sections. Gray matter signals exhibited somewhat different components than white matter signals, but both tissue types were well represented by the signal model. Frozen tissue storage did not alter the signal components but influenced component amplitudes. D2 O exchange was necessary to characterize the non-aqueous signal components, but component amplitude mapping could be reliably performed also in the presence of H2 O signals. The myelin mapping approach explored here produced reasonable and stable results for all samples. The extensive tissue and methodological investigations performed in this work form a basis for signal interpretation in future studies both ex vivo and in vivo.

First Evidence of the Expression and Localization of Prothymosin α in Human Testis and Its Involvement in Testicular Cancers.

Prothymosin α (PTMA) is a phylogenetically conserved polypeptide in male gonads of Vertebrates. In Mammals, it is a ubiquitous protein, and, possessing a random-coil structure, it interacts with many other partners, in both cytoplasmic and nuclear compartments. PTMA has been widely studied during cell progression in different types of cancer because of its anti-apoptotic and proliferative properties. Here, we provided the first evidence of PTMA expression and localization in human testis and in two testicular cancers (TC): classic seminoma (CS) and Leydig cell tumor (LCT). Data showed that its protein level, together with that of proliferating cell nuclear antigen (PCNA), a cell cycle progression marker, increased in both CS and LCT samples, as compared to non-pathological (NP) tissue. Moreover, in the two-cancer tissue, a decreased apoptotic rate and an increased autophagic flux was also evidenced. Results confirmed the anti-apoptotic action of PTMA, also suggesting that it can act as a switcher from apoptosis to autophagy, to favor the survival of testicular cancer cells when they develop in adverse environments. Finally, the combined data, even if they need to be further validated, add new insight into the role of PTMA in human normal and pathological testicular tissue.

Comparison between diffusion-weighted magnetic resonance and positron-emission tomography in the evaluation of treated lymphomas with mediastinal involvement

BackgroundThe persistence of residual tissue after treatment is frequent in patients with mediastinal lymphomas and it is often characterized by 18F-Flurodeoxyglucose Positron Emission Tomography (18F-FDG PET) uptake. This study aims to investigate the usefulness of diffusion-weighted whole-body imaging with background body signal suppression (DWIBS) sequence in residual tissues of treated mediastinal lymphomas and to compare it with 18F-FDG PET-CT.ResultsWe included 21 patients with mediastinal Hodgkin and non-Hodgkin lymphomas who showed residual masses on PET-CT imaging at end of treatment and underwent DWIBS-Magnetic Resonance Imaging (MRI). SUVmax and Apparent Diffusion Coefficient (ADC) values of residual masses were assessed quantitatively, including measurement of mean ADC. 15 patients showed radiotracer uptake at 18F-FDG PET-CT, among them only 3 had positive DWIBS-MRI with low ADC values (median value: 0.90 mm2/s). The mediastinal biopsy in these 3 “double positive” patients confirmed pathological residual tissue. All the patients with positive 18F-FDG PET-CT but negative DWIBS-MRI (n = 18) with high ADC values (median value: 2.05 mm2/s) were confirmed negative by biopsy.ConclusionsDWIBS-MRI examination combined with ADC measurement allowed to discriminate pathological and non-pathological residual tissue in patients with treated mediastinal lymphoma. These preliminary results seem to pave the way for a leading role of the MRI which could be a useful alternative to the 18F-FDG PET/CT.

Microsurgical approaches to the pulvinar: A comparative analysis

ObjectiveTo compare the area of exposure to the cisternal thalamus associated with four surgical techniques: supracerebellar-infratentorial (SCIT), occipital interhemispheric (OI), transchoroidal (TC) and subtemporal before and after parahippocampal resection (ST and STh, respectively). MethodsAll approaches were performed on both sides of three heads. Qualitative anatomical analyses were performed to understand anatomical limits, advantages, and flaws of each technique. Quantitative analyses for multiple repeated dependent variables assessed significant differences between areas of exposure. ResultsExposure area was significantly more extensive using TC and STh approaches compared to ST, OI, and SCIT. STh achieved a significantly wider exposure compared to ST. Regarding dissection angle, surrounding structures and limitations, ST approaches do not provide adequate exposure, nor alignment with the thalamic axis. The OI and STh may provide a better field of exposure, but without adequate alignment and challenging deeper dissections. TC provides better exposure of the cisternal pulvinar with access to lateral pulvinar at the atrium’s anterior wall but is a transcortical route that disrupts non-pathological tissue. SCIT provides an adequate area of exposure with the possibility of alignment with the thalamus axis, thus allowing an easier dissection of deeper lesions. ConclusionsFor lesions at the pulvinar surface, OI and STh are adequate. For lesions restricted to medial pulvinar and deep along the thalamus axis, SCIT approaches are recommended. Lesions extending to the lateral pulvinar and ventricular atrium are best removed through TC approaches. The ST approach was not suitable to the cisternal pulvinar due to its limited angular exposure.

Clival Salivary Heterotopia (Salivary Choristoma): A Rare Radiopathogical Lesión

Objective: To describe the radiological and pathological presentation of a salivary heterotopia in clivus. The lesion was detected incidentally in a 57-year-old woman. Material and Methods: Radiologic imaging (CT scan and MRI) and histological picture obtained from a clival lesion. Results: Radiologically the lesion presented as a well-limited lytic lesion with a diameter of 1.5 cm and accompanied by a cerebrospinal fluid secondary fistula. Histological examination revealed non-pathological salivary gland tissue. Conclusion: The differential diagnosis of this rare entity mandates a histological examination of the lesion.

Canonical and Non-Canonical Roles of PFKFB3 in Brain Tumors.

PFKFB3 is a bifunctional enzyme that modulates and maintains the intracellular concentrations of fructose-2,6-bisphosphate (F2,6-P2), essentially controlling the rate of glycolysis. PFKFB3 is a known activator of glycolytic rewiring in neoplastic cells, including central nervous system (CNS) neoplastic cells. The pathologic regulation of PFKFB3 is invoked via various microenvironmental stimuli and oncogenic signals. Hypoxia is a primary inducer of PFKFB3 transcription via HIF-1alpha. In addition, translational modifications of PFKFB3 are driven by various intracellular signaling pathways that allow PFKFB3 to respond to varying stimuli. PFKFB3 synthesizes F2,6P2 through the phosphorylation of F6P with a donated PO4 group from ATP and has the highest kinase activity of all PFKFB isoenzymes. The intracellular concentration of F2,6P2 in cancers is maintained primarily by PFKFB3 allowing cancer cells to evade glycolytic suppression. PFKFB3 is a primary enzyme responsible for glycolytic tumor metabolic reprogramming. PFKFB3 protein levels are significantly higher in high-grade glioma than in non-pathologic brain tissue or lower grade gliomas, but without relative upregulation of transcript levels. High PFKFB3 expression is linked to poor survival in brain tumors. Solitary or concomitant PFKFB3 inhibition has additionally shown great potential in restoring chemosensitivity and radiosensitivity in treatment-resistant brain tumors. An improved understanding of canonical and non-canonical functions of PFKFB3 could allow for the development of effective combinatorial targeted therapies for brain tumors.

Fluorescence diagnostics and photodynamic therapy of squamous cell carcinoma of the lateral surface of the tongue using the photosensitizer chlorin e6 by spectroscopic video fluorescence methods

This paper presents the results of using intraoperative fluorescence diagnostics (FD) with the endoscopic video system to increase the efficiency of photodynamic therapy (PDT). The molecular form of chlorin e6 was used as a photosensitizer (PS). All patients received an intravenous administration drug based on chlorin e6 at the concentration of 1 mg/kg. Diagnostics and therapy were carried out in 4 patients diagnosed by malignant neoplasms of the lateral surface of the tongue. Determination of the boundaries by changing the signal of the fluorescence intensity of the tumor was carried out before PDT and after PDT. The efficiency of PDT was assessed by the fluorescent signal of the tumor decreasing when compared with non-pathological normal tissue. The FD method allows to determine accurately the actual size of the tumor and its borders. All patients underwent PDT influenced by the therapeutic laser with a generation wavelength of 660 nm. It is shown that the using of fluorescence diagnostics improves the quality of the photodynamic therapy, since it is possible to assess the photobleaching of the drug during the operation.

Cellular Substrates of Functional Network Integration and Memory in Temporal Lobe Epilepsy

Temporal lobe epilepsy (TLE) patients are at risk of memory deficits, which have been linked to functional network disturbances, particularly of integration of the default mode network (DMN). However, the cellular substrates of functional network integration are unknown. We leverage a unique cross-scale dataset of drug-resistant TLE patients (n = 31), who underwent pseudo resting-state functional magnetic resonance imaging (fMRI), resting-state magnetoencephalography (MEG) and/or neuropsychological testing before neurosurgery. fMRI and MEG underwent atlas-based connectivity analyses. Functional network centrality of the lateral middle temporal gyrus, part of the DMN, was used as a measure of local network integration. Subsequently, non-pathological cortical tissue from this region was used for single cell morphological and electrophysiological patch-clamp analysis, assessing integration in terms of total dendritic length and action potential rise speed. As could be hypothesized, greater network centrality related to better memory performance. Moreover, greater network centrality correlated with more integrative properties at the cellular level across patients. We conclude that individual differences in cognitively relevant functional network integration of a DMN region are mirrored by differences in cellular integrative properties of this region in TLE patients. These findings connect previously separate scales of investigation, increasing translational insight into focal pathology and large-scale network disturbances in TLE.

Fluorescence Lifetime Imaging and Spectroscopic Co-Validation for Protoporphyrin IX-Guided Tumor Visualization in Neurosurgery

Maximal safe resection is a key strategy for improving patient prognosis in the management of brain tumors. Intraoperative fluorescence guidance has emerged as a standard in the surgery of high-grade gliomas. The administration of 5-aminolevulinic acid prior to surgery induces tumor-specific accumulation of protoporphyrin IX, which emits red fluorescence under blue-light illumination. The technology, however, is substantially limited for low-grade gliomas and weakly tumor-infiltrated brain, where low protoporphyrin IX concentrations are outweighed by tissue autofluorescence. In this context, fluorescence lifetime imaging has shown promise to distinguish spectrally overlapping fluorophores. We integrated frequency-domain fluorescence lifetime imaging in a surgical microscope and combined it with spatially registered fluorescence spectroscopy, which can be considered a research benchmark for sensitive protoporphyrin IX detection. Fluorescence lifetime maps and spectra were acquired for a representative set of fresh ex-vivo brain tumor specimens (low-grade gliomas n = 15, high-grade gliomas n = 80, meningiomas n = 41, and metastases n = 35). Combining the fluorescence lifetime with fluorescence spectra unveiled how weak protoporphyrin IX accumulations increased the lifetime respective to tissue autofluorescence. Infiltration zones (4.1ns ± 1.8ns, p = 0.017) and core tumor areas (4.8ns ± 1.3ns, p = 0.040) of low-grade gliomas were significantly distinguishable from non-pathologic tissue (1.6ns ± 0.5ns). Similarly, fluorescence lifetimes for infiltrated and reactive tissue as well as necrotic and core tumor areas were increased for high-grade gliomas and metastasis. Meningioma tumor specimens showed strongly increased lifetimes (12.2ns ± 2.5ns, p = 0.005). Our results emphasize the potential of fluorescence lifetime imaging to optimize maximal safe resection in brain tumors in future and highlight its potential toward clinical translation.

Preliminary Investigation on the Involvement of Cytoskeleton-Related Proteins, DAAM1 and PREP, in Human Testicular Disorders.

Herein, for the first time, the potential relationships between the cytoskeleton-associated proteins DAAM1 and PREP with different testicular disorders, such as classic seminoma (CS), Leydig cell tumor (LCT), and Sertoli cell-only syndrome (SOS), were evaluated. Six CS, two LCT, and two SOS tissue samples were obtained during inguinal exploration in patients with a suspect testis tumor based on clinical examination and ultrasonography. DAAM1 and PREP protein levels and immunofluorescent localization were analyzed. An increased DAAM1 protein level in CS and SOS as compared to non-pathological (NP) tissue was observed, while LCT showed no significant differences. Conversely, PREP protein level increased in LCT, while it decreased in CS and SOS compared to NP tissue. These results were strongly supported by the immunofluorescence staining, revealing an altered localization and signal intensity of DAAM1 and PREP in the analyzed samples, highlighting a perturbed cytoarchitecture. Interestingly, in LCT spermatogonia, a specific DAAM1 nuclear localization was found, probably due to an enhanced testosterone production, as confirmed by the increased protein levels of steroidogenic enzymes. Finally, although further studies are needed to verify the involvement of other formins and microtubule-associated proteins, this report raised the opportunity to indicate DAAM1 and PREP as new potential markers, supporting the cytoskeleton dynamics changes occurring during normal and/or pathological cell differentiation.

The Gene Expression of MMP-2 and TIMP-1 in Non-pathological Paratumoral and Autopsy Lung Tissues

Background: Finding a sample of healthy tissue is a critical challenge in research studies. Non-pathological Tissue adjacent to the tumor (NAT) specimens is usually used as the control in several studies. However, little is known about the similarity of NAT to healthy tissues. Here, we compared the expression of Matrix Metalloproteinase 2 (MMP-2) and its inhibitor, Tissue Inhibitors of MMP (TIMP)-1 as extracellular matrix remodeling factors in NAT and autopsy lung tissue. Materials and Methods: RNA of 7 NAT and 6 Formalin-Fixed Paraffin-Embedded (FFPE) lung autopsies from healthy people as the control group was extracted, and cDNA was synthesized. The gene expression levels of MMP-2 and TIMP-1 were evaluated by real-time PCR. Results: There were no significant differences in the expression of MMP-2, TIMP-1, or their ratio between the two groups. Conclusion: The results showed that NAT could be used as healthy controls in lung tissue studies for MMP-2 and TIMP-1.

Convergent and distributed effects of the 3q29 deletion on the human neural transcriptome

The 3q29 deletion (3q29Del) confers high risk for schizophrenia and other neurodevelopmental and psychiatric disorders. However, no single gene in this interval is definitively associated with disease, prompting the hypothesis that neuropsychiatric sequelae emerge upon loss of multiple functionally-connected genes. 3q29 genes are unevenly annotated and the impact of 3q29Del on the human neural transcriptome is unknown. To systematically formulate unbiased hypotheses about molecular mechanisms linking 3q29Del to neuropsychiatric illness, we conducted a systems-level network analysis of the non-pathological adult human cortical transcriptome and generated evidence-based predictions that relate 3q29 genes to novel functions and disease associations. The 21 protein-coding genes located in the interval segregated into seven clusters of highly co-expressed genes, demonstrating both convergent and distributed effects of 3q29Del across the interrogated transcriptomic landscape. Pathway analysis of these clusters indicated involvement in nervous-system functions, including synaptic signaling and organization, as well as core cellular functions, including transcriptional regulation, posttranslational modifications, chromatin remodeling, and mitochondrial metabolism. Top network-neighbors of 3q29 genes showed significant overlap with known schizophrenia, autism, and intellectual disability-risk genes, suggesting that 3q29Del biology is relevant to idiopathic disease. Leveraging “guilt by association”, we propose nine 3q29 genes, including one hub gene, as prioritized drivers of neuropsychiatric risk. These results provide testable hypotheses for experimental analysis on causal drivers and mechanisms of the largest known genetic risk factor for schizophrenia and highlight the study of normal function in non-pathological postmortem tissue to further our understanding of psychiatric genetics, especially for rare syndromes like 3q29Del, where access to neural tissue from carriers is unavailable or limited.

Overexpression of PELP1 in Lung Adenocarcinoma Promoted E2 Induced Proliferation, Migration and Invasion of the Tumor Cells and Predicted a Worse Outcome of the Patients.

The expression of Proline-, glutamic acid-, and leucine-rich protein 1 (PELP1) has been reported to be dysregulated in non-small cell lung carcinoma, especially in lung adenocarcinoma (LUAD). Therefore, we aimed to investigate the functional and prognostic roles of PELP1 in LUAD in this study. We first immunolocalized PELP1 in 76 cases of LUAD and 17 non-pathological or tumorous lung (NTL) tissue specimens and correlated the findings with the clinicopathological parameters of the patients. We then performed in vitro analysis including MTT, flow cytometry, wound healing, and transwell assays in order to further explore the biological roles of PELP1 in 17-β-estradiol (E2) induced cell proliferation, migration, and invasion of LUAD cells. We subsequently evaluated the prognostic significance of PELP1 in LUAD patients using the online survival analysis tool Kaplan-Meier Plotter. The status of PELP1 immunoreactivity in LUAD was significantly higher than that in the NTL tissues and significantly positively correlated with less differentiated features of carcinoma cells, positive lymph node metastasis, higher clinical stage as well as the status of ERα, ERβ, and PCNA. In vitro study did reveal that E2 promoted cell proliferation and migration and elevated PELP1 protein level in PELP1-high A549 and H1975 cells but not in PELP1-low H-1299 cells. Knock down of PELP1 significantly attenuated E2 induced cell proliferation, colony formation, cell cycle progress as well as migration and invasion of A549 and H1975 cells. Kaplan-Meier Plotter revealed that LUAD cases harboring higher PELP1 expression had significantly shorter overall survival. In summary, PELP1 played a pivotal role in the estrogen-induced aggressive transformation of LUAD and could represent adverse clinical outcome of the LUAD patients.