Stromal Fibrosis Research Articles

Abstract CT549: Randomized Phase II trial of two different nutritional approaches for patients receiving treatment for their advanced pancreatic cancer

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) is characterized by stromal fibrosis, hypoxia, and nutritional deprivation. PDAC tumors grow aggressively, diagnosis is typically made after metastasis and the disease remains associated with poor outcomes. The triplet chemotherapy regimen of gemcitabine, nab-paclitaxel with cisplatin was associated with a median overall survival of 16.4 months in patients with metastatic pancreatic cancer in the first-line setting (Jameson et al., 2020). Nutritional, metabolic interventions offer an opportunity to fundamentally change the tumor microenvironment and improve outcomes for patients. A medically supervised ketogenic diet (MSKD) defined as lower carbohydrate, lower protein, and higher fat can significantly reduce glucose and insulin and increase metabolically active ketone bodies. A ketogenic diet combined with triplet chemotherapy (gemcitabine, nab-paclitaxel, cisplatin) was shown to inhibit murine pancreatic KPC tumor growth and to triple the survival benefit of chemotherapy alone. The ketogenic diet combined with triple chemotherapy was associated with glucose depletion, altered TCA substrate usage, and NADH elevation. Methods: In this Phase II randomized clinical trial (NCT04631445), we are evaluating a medically supervised ketogenic diet (MSKD) versus a standard diet when combined with the triplet therapy in patients with treatment-naive advanced pancreatic cancer. The primary endpoint is progression free survival for triplet therapy while on MSKD or non-MSKD. Secondary endpoints include disease control rate (PR+ CR+ SD for at least 9 weeks), change in CA 19-9 (or CA125, or CEA if not expressers of CA 19-9), average insulin levels, HbA1c, body weight, a comparison of gut microbial diversity, changes in serum metabolites and quality of life via the EORTC QLQ-C30 assessment. Unlike prior ketogenic intervention studies, the MSKD is being supported by a continuous care nutrition intervention through Virta Health Corp, that offers tracking of daily ketone and glucose levels, a web-based software application, education, and communication with a remote care team to ensure sustained nutritional ketosis. A total of 40 patients with untreated metastatic PDAC are planned for enrollment, 20 randomized to each arm. The trial opened for accrual November 2020. Citation Format: Diana Hanna, Gayle S. Jameson, Drew W. Rasco, Angela Alistar, Richard C. Frank, Anthony B. El-Khoueiry, Julia E. Wiedmeier, Caroline Roberts, Brandon Fell, Sarah Hallberg, Denise Roe, Derek Cridebring, Joshua Rabinowitz, Stephen Thomas Gately, Daniel D. Von Hoff. Randomized Phase II trial of two different nutritional approaches for patients receiving treatment for their advanced pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT549.

Abstract 935: NEK1 phosphorylates YAP1 to promote prostate cancer tumorigenesis

Abstract Background: Prostate cancer (PCa) is the second leading cause of cancer death in men of western part of the world. The disease progression has been defined by substages (adenocarcinoma to castration resistant PCa to neuroendocrine PCa) both from mouse models and patients from clinical studies. The mechanism of the disease advancement and hence treatment modalities is still under investigation. Hippo pathway, the evolutionarily conserved developmental pathway, known to regulate organ size, proliferation, apoptosis, migration, stemness etc., is also implicated in PCa tumorigenesis. Transcriptional co-activator YAP1 which acts downstream of the canonical hippo kinases is known to interact with plethora of proteins. But how non-canonical hippo kinases can regulate YAP1 in this disease is yet to be unraveled. We find that NIMA-related kinase isoform 1, Nek1, interacts with YAP1 and phosphorylates Tyr-407 of YAP1. The significance of this post-translational modification is the focus of the study. We find that Nek1 phosphorylates YAP1 to stabilize YAP1 and promote tumorigenesis. Material and Methods: Human YAP1 cDNA encoding plasmid mutated at Tyr 407 to Phenylalanine using site-directed mutagenesis. YAP1-Y407F and WT expressed in HEK293A cells. For genetic analysis in mouse model, female TRAMP mice were bred with Nek1 -/+ male mouse (Jackson laboratory). Genotypic analysis confirmed Nek1-/+ TRAMP and Nek1+/+TRAMP which were used for this study. After 16weeks of age the male mice were castrated and sacrificed at 20weeks of age. Prostate tissues collected and histological sections done (paraffin embedded). Immunohistochemical (IHC) analysis was done using vector labs IHC kits, data analyzed and scored by our pathologist. Results: Immunoblots from YAP1-Y407F and WT cells treated with Cycloheximide (50ug/ml) at different time points (0.5-6hrs) show that YAP1-Y407F is less stable than WT with t1/2 of 0.5hrs whereas WT is stable till 2hrs. Very importantly, while intact TRAMP/Nek1+/- mice do develop PCa normally with much enlargement of the prostate at 18-20 weeks, if these mice are castrated, the tumors fail to grow and the prostate retains normal size and general morphology. Our IHC data confirms that firstly, castrated Nek1+/+TRAMP shows clearly cancerous acini whereas the Nek-/+ TRAMP shows mostly stromal fibrosis and hyperplasia (as shown in H&E and IHC). Secondly, when stained for activated Nek1 marker, pNek1 (T141), we find that Nek1 stain intensity is low in Nek1-/+ tissues compared to Nek1+/+. Most importantly, YAP1 level is lower in the same parallel tissue sections of Nek1-/+. From the above analysis, we conclude that Nek1 activity regulates stability of YAP1, and infer that YAP1 function is critical for the timely progression to adenocarcinoma following androgen deprivation therapy. Acknowledgements: Funding Resources: Department of Defense (DoD-PCRP grant W81XWH-17-1-0417) ; FWCC Bridge Award. Citation Format: Ishita Ghosh, Zobair Alam, Md. Imtiaz Khalil, Judy King, Arrigo DeBenedetti. NEK1 phosphorylates YAP1 to promote prostate cancer tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 935.

Characteristics of the reproductive mucosa microbiota in patients with endometrial hyperplasia in combination with chronic endometritis and an optimized program of rehabilitation therapy and preconception training

The etiology and pathogenesis of hyperplastic processes of the endometrium (HPE) are currently unclear, some aspects of the pathogenesis remain controversial and debatable. The objective: was to evaluate the microbiota of the reproductive mucosa in patients with subfertility on the background of endometrial hyperplasia combined with chronic endometritis, and to develop an optimized treatment and rehabilitation program. Materials and methods. A comprehensive clinical and laboratory examination of 54 patients of reproductive age with subfertility on the background of endometrial hyperplasia and chronic endometritis was performed. Two groups were formed to evaluate the effectiveness: the main group - 30 patients who received an optimized program of treatment and rehabilitation options, the comparison group – 24 patients who received classic conventional therapy. The endometrium of 20 gynecologically healthy women was used for control clinical-laboratory and morphological examinations. The personalized program of postoperative support is offered: basic antibiotic therapy, supplemented with bacteriophage therapy, antiviral therapy, immunomodulatory, angioprotective and venotonizing agents, correction of ischemia, correction of metabolic disorders, restoration of morphofunctional potential of endometrial tissue with the use of individualized hormone therapy was proposed. Results. HPE became the prerogative of all patients in the study group, and in 19 (35.2%) endometrial polyps were registered, in 35 (64.8%) a combination of hyperplasia and polyps. The results of bacteriological research showed a significant proportion of abnormalities in vaginal biocinosis, which is manifested by a significant reduction in the frequency of secretions of major acid-forming lactobacilli compared with healthy patients. After using the proposed treatment program to improve well-being, reduce pain and reduce hypersecretory abnormalities were noted by all patients, the results of control pipelle biopsy with microbiological examination of the endometrium 3 months after the end of treatment showed sterile cultures in 97% of samples. During the year of observation, spontaneous pregnancy out of 30 people who received pre-pregnancy training in the main group occurred in 14 (46.7% of cases), and of the 16 women enrolled in the ART program, 7 (43.7%) became pregnant. Conclusions. A comprehensive study of the microbiocenosis of the mucous membranes of the reproductive tract showed that in women with endometrial hyperplasia combined with CE, microbiological parameters are characterized by a high frequency of viral and viral-bacterial associations, a significant proportion of dysbiotic disorders. Carrying out pregravid preparation along with optimizing the antibacterial therapy program using rational antimicrobial agents, modern enzyme preparations to reduce the intensity of stroma fibrosis, individualized hormonal correction, agents that improve the effects of ischemia and changes in mucosal microbiota, allow to restore morphofunctional disorders in the endometrium.

Primary malignant melanoma of the esophagus: differentiation from esophageal squamous cell carcinoma and leiomyoma using dynamic contrast-enhanced CT findings

PurposeThis study aimed to summarize the computed tomography (CT) findings of PMME and differentiate it from esophageal SCC and leiomyoma using CT analysis.MethodsThis was a retrospective study including 23 patients with PMME, 69 patients with SCC, and 21 patients with leiomyoma in our hospital. Qualitative CT morphological characteristics of each lesion included the location, tumor range, ulcer, enhanced pattern, and so on. For quantitative CT analysis, thickness, length and area of tumor, size of largest lymph node, number of metastatic lymph node, and CT value of tumor in plain, arterial, and delayed phases were measured. The associated factors for differentiating PMME from SCC and leiomyoma were examined with univariate and multivariate analysis. Receive operating characteristic curve (ROC) was used to determine the performance of CT models in discriminating PMME from SCC and leiomyoma.ResultsThe thickness, mean CT value in arterial phase, and range of tumor were the independent factors for diagnosing PMME from SCC. These parameters were used to establish a diagnostic CT model with area under the ROC (AUC) of 0.969, and accuracy of 90.2%. In pathology, interstitial vessels in PMME were more abundant than that of SCC, and the stromal fibrosis was more obvious in SCC. PMME commonly exhibited intraluminal expansively growth pattern and SCC often showed infiltrative pattern. The postcontrast attenuation difference in maximum CT attenuation value between plain and arterial phases was the independent factor for diagnosing PMME from leiomyoma. This parameter was applied to differentiate PMME from leiomyoma with AUC of 0.929 and accuracy of 86.4%.ConclusionThe qualitative and quantitative CT analysis had excellent performance for differentiating PMME from SCC and esophageal leiomyoma.Graphical abstract

Estrogen and G protein-coupled estrogen receptor accelerate the progression of benign prostatic hyperplasia by inducing prostatic fibrosis

Benign prostatic hyperplasia (BPH) is the most common and progressive urological disease in elderly men worldwide. Epidemiological studies have suggested that the speed of disease progression varies among individuals, while the pathophysiological mechanisms of accelerated clinical progression in some BPH patients remain to be elucidated. In this study, we defined patients with BPH as belonging to the accelerated progressive group (transurethral resection of the prostate [TURP] surgery at ≤50 years old), normal-speed progressive group (TURP surgery at ≥70 years old), or non-progressive group (age ≤50 years old without BPH-related surgery). We enrolled prostate specimens from the three groups of patients and compared these tissues to determine the histopathological characteristics and molecular mechanisms underlying BPH patients with accelerated progression. We found that the main histopathological characteristics of accelerated progressive BPH tissues were increased stromal components and prostatic fibrosis, which were accompanied by higher myofibroblast accumulation and collagen deposition. Mechanism dissection demonstrated that these accelerated progressive BPH tissues have higher expression of the CYP19 and G protein-coupled estrogen receptor (GPER) with higher estrogen biosynthesis. Estrogen functions via GPER/Gαi signaling to modulate the EGFR/ERK and HIF-1α/TGF-β1 signaling to increase prostatic stromal cell proliferation and prostatic stromal fibrosis. The increased stromal components and prostatic fibrosis may accelerate the clinical progression of BPH. Targeting this newly identified CYP19/estrogen/GPER/Gαi signaling axis may facilitate the development of novel personalized therapeutics to better suppress the progression of BPH.

Application of deep learning models on whole slide images uncover new histological markers related to high-risk malignant pleural mesothelioma.

e13580 Background: Malignant Pleural mesothelioma (MPM) is a highly aggressive cancer of the pleural surface and represents 80-90% of mesothelioma diagnosis. MPM is broadly subclassified into three histological subgroups (epithelioid, sarcomatoid, biphasic), however tissue heterogeneity has resulted in diagnostic challenges and suboptimal patient care. There are currently no specific histological markers of high/low-risk MPM patients, which is critical in predicting patient prognosis. Methods: Owkin developed MesoNet, a deep learning model that predicts overall survival (OS) of MPM patients from whole slide images (WSI) and trained on the French MESOBANK and TCGA cohorts (Courtiol et al, 2019). In this study, we sought to validate MesoNet’s performance on an independent cohort of 127 WSI stained with haematoxylin/eosin from MPM patients collected at the University of Pittsburgh as part of the National Mesothelioma Virtual Bank (funding by U24OH009077). Patient demographics, survival data, expertly curated pathology annotations were also collected. Results: Our analyses showed that MesoNet predicted OS as risk score based on WSI, which validated high-risk MPM patients exhibited poorer OS, as compared to low-risk patients. Analyses on histological subtypes revealed sarcomatoid and biphasic patients were overrepresented in high-risk groups, as compared to epithelioid patients, which correlates with observed OS data. Notably, histological features associated with high-risk patients revealed tumor pleomorphism and anaplastic nuclear features, whereas low-risk tiles appear to be enriched in tumor infiltrating lymphocytes (TILs) with accompanying stromal proliferation and dense fibrosis. Conclusions: Collectively, our studies validate MesoNet performance on an independent cohort and identify new features related to MPM risk groups, which may inform future treatment stratification and personalization of immunotherapies.

Phase III study (daNIS-2) of the anti–TGF-β monoclonal antibody (mAb) NIS793 with nab-paclitaxel/gemcitabine (NG) versus NG alone in patients (pts) with first-line metastatic pancreatic ductal adenocarcinoma (mPDAC).

TPS4193 Background: Despite improving outcomes, current therapies for mPDAC have a modest impact on overall survival (OS) and new therapies are needed. PDAC is characterized by an abundance of intratumoral fibrosis, which may contribute to the lack of treatment efficacy and act as a mechanical barrier to effective penetration of therapeutics. TGF-β has a multifactorial role in tumorigenesis and maintaining an immunosuppressive tumor microenvironment (TME). Emerging evidence points to the role of TGF-β as a pivotal activator of cancer-associated fibroblasts that lead to the development of fibrotic networks. In preclinical models, TGF-β blockade alters the TME to facilitate an antitumor response, reduce stromal fibrosis, and augment the benefit of chemotherapy, providing rationale for combining TGF-β–targeting agents with chemotherapy. NIS793 is a potent, selective, human IgG2 mAb antagonist of TGF-β. This study investigates NIS793 in combination with NG vs NG alone in treatment-naïve pts with mPDAC. Methods: This is a phase III, randomized, double-blind, multicenter, two-arm study (NCT04935359) consisting of two stages: an initial safety run-in period followed by two-arm randomization. Eligible pts include adults with previously untreated mPDAC and an ECOG performance status ≤1. Pts with a tumor histology other than adenocarcinoma or with microsatellite instability-high tumors are ineligible. The aim of the safety run-in period is to assess the safety and tolerability of NIS793 + NG and confirm the recommended dose for the randomized phase of this study. Data will be analyzed once at least six evaluable pts have received NIS793 (intravenous [IV] 2100 mg every 2 weeks) + nab-paclitaxel (IV 125 mg/m2 on Days 1, 8, and 15) + gemcitabine (IV 1000 mg/m2 on Days 1, 8, and 15) for one 28-day cycle. Pts (N = 480) will be randomized 1:1 to NIS793 + NG or placebo + NG. Treatment will continue until unacceptable toxicity, disease progression, discontinuation by investigator or pt choice, death, or withdrawal of consent. The primary objective is to evaluate the OS of pts receiving NIS793 + NG vs NG alone; secondary objectives include assessing progression-free survival, the overall response rate, disease control rate, duration of response, and time to response (assessed locally per RECIST v1.1), as well as safety and tolerability, immunogenicity, pharmacokinetics, and patient-reported outcomes such as health-related quality of life. Efficacy will be assessed at screening, every 8 weeks for 1 year, and then every 12 weeks until disease progression. Blood samples will be taken at baseline and during treatment for pharmacokinetic and immunogenicity assessments. This study is ongoing and will enroll pts from approximately 149 sites across 28 countries. The first pt was treated on October 20, 2021. Clinical trial information: NCT04935359.

Immunomorphological signs of chronic endometritis in patients after failed attempts of auxiliary reproductive technologies

The objective: to analyze the morphological and immunohistochemical features of the endometrium in the patients with chronic endometritis after unsuccessful attempts of assisted reproductive technologies (ART). Materials and methods. A prospective study was conducted in women with a history of chronic endometritis on the basis of the Medical Center LLC “ISIDA-IVF” for the period from 2019 to 2021. The I group included 55 women of reproductive age with chronic endometritis and repeated unsuccessful attempts of ART, the control group (CG) – 40 women of reproductive age without fertility disorders who are preparing for the replacement motherhood programs or who are oocyte donors. The results of the endometrial biopsy performed on the 5–7th and 19th–21st days of the menstrual cycle using a pipelle aspiration curette were studied for the assessment of the immunomorphological characteristics of the endometrium in the patients with chronic endometritis. The immunoperoxidase method with primary specific monoclonal antibodies was used for the immunomorphological study of estrogen and progesterone receptors. Differences at p<0.05 were considered statistically significant. Results. Pathomorphological study of the endometrium revealed characteristic signs of chronic inflammation – the presence of a large number of plasma cells in the endometrium stroma, lymphoid infiltrates near the blood vessels and the gland orifices, unevenly expressed stroma fibrosis, sclerotic changes in the walls of spiral arteries. All examined persons had two or more morphological signs of chronic endometritis (CE). An increase number of macrophages and monocytes in the endometrium glands and stroma was found by CE – 31.3±2.9 % versus 2.3±0.1 % in CG (р<0.05). An increase proliferative activity of epithelium cells of the endometrium glands and stroma on the 5-7th day of the menstrual cycle in patients in the I group compared to similar indicators of the secretion phase in CG was determined. The proliferation of glandular and stromal cells, detected on the 19th–21st days of the menstrual cycle, had the intensification of the programmed cell death process in the covering and glandular epithelium of the endometrium. This was characterized by the structural changes in the cell nuclei – the marginal state of chromatin, the appearance of karyorrhexis and karyopyknosis. In the endometrium stroma the presence of capillary-type vessels was found, the intensity of their development had a strong correlation with the degree of severity of the inflammatory changes (r=0.74; p<0.05). During the study of progesterone receptors mainly a decrease of the level of expression in the glands both compared to the indicators of healthy women and in the secretion phase compared to the proliferation phase - H-score 145.5±7.7 was found, in the proliferation phase versus H-score 55.5±10.4 in the secretion phase (р<0.05). Conclusions. Immunomorphological study of the endometrium in patients with chronic endometritis after unsuccessful attempts of assisted reproductive technologies demonstrates that the inflammatory changes in the tissues have clear morphological criteria, which are most pronounced on the 5-7th day of the menstrual cycle – the presence of plasma cells, stroma fibrosis, sclerosis vessel walls, decrease of the estrogen and progesterone level receptors. These morphological and immunohistochemical changes persist in the secretory phase of the menstrual cycle with the most significant manifestations in the form of a decrease in the level of progesterone receptors.

Umbilical cord mesenchymal stem cell-derived exosomes reverse endometrial fibrosis by the miR-145-5p/ZEB2 axis in intrauterine adhesions

Research questionWhat is the specific mechanism of umbilical cord mesenchymal stem cell-derived exosomes (UCMSC-exos) in regulating endometrial repair and regeneration? DesignIn this study, UCMSC-exos were harvested by differential ultracentrifugation from umbilical cord mesenchymal stem cell culture supernatant and identified with western blotting, transmission electron microscopy and nanoparticle tracking analysis. Transforming growth factor-β1 (TGFβ1) at different concentrations was used to construct the intrauterine adhesions cell model. The fibrotic markers were assessed by quantitative reverse transcription-polymerase chain reaction and western blotting. The effects of miR-145-5p over-expression on endometrial fibrosis were assessed. Dual luciferase assay was performed to verify the relationship between miR-145-5p and zinc finger E-box binding homeobox 2 (ZEB2). ResultsThe isolated UCMSC-exos had a typical cup-shaped morphology, expressed the specific exosomal markers Alix, CD63 and TSG101, and were approximately 50–150 nm in diameter. TGFβ1 at 10 ng/ml significantly promoted endometrial fibrosis, which was reversed by 20 µg/ml UCMSC-exos. Exosomal miR-145-5p ameliorated TGFβ1-induced endometrial fibrosis. ZEB2 was inversely regulated by exosomal miR-145-5p as a direct target. ConclusionsUCMSC-exos might reverse endometrial stromal cell fibrosis by regulating the miR-145-5p/ZEB2 axis, representing a potential novel strategy to promote endometrial repair.

Trophic and immunomodulatory effects of adipose tissue derived stem cells in a preclinical murine model of endometriosis

Endometriosis, which exhibits enigmatic pathological features such as stromal fibrosis and proliferation of ectopic epithelial cells, is known as a refractory disease. Mesenchymal stem cells modulate the fibrosis in stromal tissues through their trophic and immunomodulatory properties. To investigate the potential of stem cells in treating endometriosis, we examined the secondary morphology and molecular alterations in endometriosis-like lesions after the administration of adipose tissue-derived stem cells (ASCs) to an experimental murine model of endometriosis. The infused ASCs were found integrated in the endometriosis-like lesions. Accompanied by the suppression of stromal fibrosis and proliferation of endometriotic epithelial cells, the infusion of ASCs with stemness potential (early passage of ASCs) suppressed the growth of endometriosis-like lesions and inhibited the expression of pro-inflammatory and pro-fibrotic cytokines, whereas no significant attenuation of endometriosis-like lesions occurred after the infusion of ASCs without stemness potential (late passage of ASCs). Accordingly, the trophic and immunomodulatory properties of ASCs may regulate fibrosis in endometriosis-like lesions, suggesting that regenerative medicine could be recognized as an innovative treatment for patients with endometriosis through the accumulation of evidence of preclinical efficacy.

RNA bisulfite sequencing reveals NSUN2-mediated suppression of epithelial differentiation in pancreatic cancer.

Posttranscriptional modifications in RNA have been considered to contribute to disease pathogenesis and tumor progression. NOL1/NOP2/Sun domain family member 2 (NSUN2) is an RNA methyltransferase that promotes tumor progression in several cancers. Pancreatic cancer relapse inevitably occurs even in cases where primary tumors have been successfully treated. Associations of cancer progression due to reprogramming of the cancer methyl-metabolome and the cancer genome have been noted, but the effect of base modifications, namely 5-methylcytosine (m5C), in the transcriptome remains unclear. Aberrant regulation of 5-methylcytosine turnover in cancer may affect posttranscriptional modifications in coding and noncoding RNAs in disease pathogenesis. Mutations in NSUN2 have been reported as drivers of neurodevelopmental disorders in mice, and upregulated expression of NSUN2 in tumors of the breast, bladder, and pancreas has been reported. In this study, we conducted mRNA whole transcriptomic bisulfite sequencing to categorize NSUN2 target sites in the mRNA of human pancreatic cancer cells. We identified a total of 2829 frequent m5C sites in mRNA from pancreatic cancer cells. A total of 90.9% (2572/2829) of these m5C sites were mapped to annotated genes in autosomes and sex chromosomes X and Y. Immunohistochemistry staining confirmed that the NSUN2 expression was significantly upregulated in cancer lesions in the LSL-KrasG12D/+;Trp53fl/fl;Pdx1-Cre (KPC) spontaneous pancreatic cancer mouse model induced by Pdx1-driven Cre/lox system expressing mutant KrasG12D and p53 deletion. The in vitro phenotypic analysis of NSUN2 knockdown showed mild effects on pancreatic cancer cell 2D/3D growth, morphology and gemcitabine sensitivity in the early phase of tumorigenesis, but cumulative changes after multiple cell doubling passages over time were required for these mutations to accumulate. Syngeneic transplantation of NSUN2-knockdown KPC cells via subcutaneous injection showed decreased stromal fibrosis and restored differentiation of ductal epithelium in vivo. SIGNIFICANCE: Transcriptome-wide mRNA bisulfite sequencing identified candidate m5C sites of mRNAs in human pancreatic cancer cells. NSUN2-mediated m5C mRNA metabolism was observed in a mouse model of pancreatic cancer. NSUN2 regulates cancer progression and epithelial differentiation via mRNA methylation.

Histology of Placenta in Normotensive and Hypertensive Cases in Khyber Pakhtunkhwa: A Comparative Study

One of the most common pregnancy complications is Hypertension. It has a major effect on maternal and neonatal morbidities and fatalities. The placenta is an important organ for the maintenance of pregnancy and the nourishment of the fetus. Objective: To compare the morbid variations in the placenta of hypertensive and normotensive mothers. Methods: From the obstetrics and gynecology department, a hundred placentae of hypertension and normotensive females were collected. The placentae were preserved in formalin and sectioned. Blocks prepared were stained with hematoxylin and eosin. Results: The placentae from 50 normotensive mothers with 26±3.1 years (mean age) and 50 hypertensive mothers with 26± 2 years (mean age). In normotensive patients 12% Infarction was seen and 70% of severe hypertensive patients while syncytial knots were found in 70% of cases of severe hypertension, while calcification was observed in 90%. In severe hypertension, stroma fibrosis and hypervascularity increase in proportion to the degree of hypertension, with 50% and 30% cases seen in severe hypertension, respectively. Conclusions: In pregnancy hypertension causes, Stromal fibrosis syncytial knots, calcified, hypervascularity, all of which impair the uteroplacental flow of blood and lower newborn weight.

Previously Unreported Salivary Heterotopia of the TMJ – 23 Serous Choristomas of One Joint

<h3>Introduction</h3> Salivary gland choristomas of the soft tissues of the TMJs, independent of adjacent parotid glands, have not been previously reported. <h3>Objective</h3> To report a massive example of serous gland heterotopia of the TMJ soft tissues. Methods and Msaterials. 116 consecutive TMJ surgical samples from 57 subjects were microscopically reviewed for abnormalities. Of these, 27 joints contained previously undescribed heterotopic glandular lobules of parotid-like tissue (plus 2 mucus lobules), with no connection to parotid glands. The case of a 41 year of female with many years of joint dysfunction and severe pain was selected for specific description because she presented the most extreme example of this phenomenon. <h3>Results</h3> Two clusters comprising a total of 57 distinct lobules of mostly normal serous acinar tissue and ductal structures were embedded in or attached to the posterior-lateral ligament of the subject's left TMJ. The ligament itself was largely replaced by dense fibrous scar, with an embedded traumatic neuroma and an hypertrophied artery, while fossa curettings showed osteoarthritis. The salivary lobules appeared to represent 23 different "glands" measuring altogether 2.5 × 2.5 × 0.5 cm. Glandular parenchyma was histologically normal and mature serous tissue, except for 6 glands which showed almost complete oncocytic change, 7 showing focal regions of oncocytic change, and 2 showing acinar atrophy with stromal fibrosis and small numbers of lymphocytes (chronic sialadenitis). No large or secretory ducts were found. Serial sectioning confirmed that there was no connection to normal parotid gland. <h3>Conclusion</h3> We report a massive cluster of 23 serous gland choristomas within and attached to the posterolateral ligament of the left TMJ in a single individual. The ligament was severely scarred and some choristomas showed chronic sialadenitis, presumably from being repeatedly pressed into the joint spaces. Heterotopic serous tissue has not been previously reported from the TMJs.

Collagen Deposition Reduction by Vitamin E in Alcoholic Liver Injury

Background: Alcohol encourages fibrotic changes in liver by initiating collagen deposition in liver parenchyma. Vitamin E can slow down progress of fibrosis in alcoholic liver injury and play a defensive part against these changes. Aim To observe difference in fibrotic changes in livers of vitamin E treated and non-treated rabbits exposed to intra-gastric administration of alcohol. Study Design: Analytical experimental study Place and duration of study: Department of Anatomy, Peshawar Medical College, Peshawar Pakistan from 1st May 2019 to 30th October 2019. Methodology: Eighteen domestic type male rabbits (Oryctolaguscuniculus) were included. All animals were divided in two categories on the basis of experiment time frame. “Category E8” animals having 8 weeks’ time period and “Category E4” animals having 4 weeks experimental duration. Each category was then clustered in “Control Group A” (appropriate food as per standard laboratory requirements, and normal saline as drinking water was provided daily), “Experimental Group B” (proper standard nutrition, 30% solution of ethanol made in distal water in the dosage of 30ml per kg/day through nasogastric tube and normal saline for drinking purpose were provided daily) and “Experimental Group C” (treated with suitable standard required diet, 30% of ethanol solution made in distal water in the dosage of 30ml per kg/day via NG tube and “Vitamin E” 50mg dissolved in 2ml distal water per kg/day with same nasogastric tube daily and normal saline for drinking purpose were given). Results: Collagen fibers deposition (fibrosis) in liver stroma was detected and recorded according to Knodel histological activity index. Quantitative difference in the collagen deposition among all groups from both “Category E4” and “Category E8” presented vastly significant “P” values. Conclusion: Alcohol can cause major fibrotic changes in “liver” even if it is consumed for the minor duration of 4 to 8 weeks, and these alterations can be minimised by the proper use of vitamin e through its “antioxidant effect”. Keywords: Alcohol, Liver, Fibrosis, Vitamin E, Hepatocyte, Collagen

IRAK4 Signaling Drives Resistance to Checkpoint Immunotherapy in Pancreatic Ductal Adenocarcinoma

Checkpoint immunotherapy is largely ineffective in pancreatic ductal adenocarcinoma (PDAC). The innate immune nuclear factor (NF)-κB pathway promotes PDAC cell survival and stromal fibrosis, and is driven by Interleukin-1 Receptor Associated Kinase-4 (IRAK4), but its impact on tumor immunity has not been directly investigated. We interrogated The Cancer Genome Atlas data to identify the correlation between NF-κB and T cell signature, and a PDAC tissue microarray (TMA) to correlate IRAK4 activity with CD8+ T cell abundance. We performed RNA sequencing (RNA-seq) on IRAK4-deleted PDAC cells, and single-cell RNA-seq on autochthonous KPC (p48-Cre/TP53f/f/LSL-KRASG12D) mice treated with an IRAK4 inhibitor. We generated conditional IRAK4-deleted KPC mice and complementarily used IRAK4 inhibitors to determine the impact of IRAK4 on T cell immunity. We found positive correlation between NF-κB activity, IRAK4 and T cell exhaustion from The Cancer Genome Atlas. We observed inverse correlation between phosphorylated IRAK4 and CD8+ T cell abundance in a PDAC tissue microarray. Loss of IRAK4 abrogates NF-κB activity, several immunosuppressive factors, checkpoint ligands, and hyaluronan synthase 2, all of which drive T cell dysfunction. Accordingly, conditional deletion or pharmacologic inhibition of IRAK4 markedly decreased tumor desmoplasia and increased the abundance and activity of infiltrative CD4+ and CD8+ T cells in KPC tumors. Single-cell RNA-seq showed myeloid and fibroblast reprogramming toward acute inflammatory responses following IRAK4 inhibition. These changes set the stage for successful combination of IRAK4 inhibitors with checkpoint immunotherapy, resulting in excellent tumor control and markedly prolonged survival of KPC mice. IRAK4 drives T cell dysfunction in PDAC and is a novel, promising immunotherapeutic target.

Clinicopathological study and immunohistochemical analysis of expression of annexin A5 and apelin in human placentae of gestational diabetes mellitus

Background and aimsGestational diabetes mellitus (GDM) is one of the commonest medical complications of pregnancy. Annexin A5 (ANXA5) is a protein, found in apical surfaces of syncytiotrophoblasts, which prevents fetal and placental vascular thrombosis in GDM. Apelin is a bioactive peptide which has been linked to GDM. The aim of the present study was to correlate macroscopic as well as microscopic changes and immunohistochemical expression of ANXA5 and apelin in placentae of GDM with maternal and neonatal clinical features and also to compare the results with those in matched controls. MethodsThis prospective observational study was undertaken for a period of one year from April 2020 to March 2021. It comprised of 42 patients of GDM. Gross features, microscopic features and intensity and grade of expression of ANXA5 and Apelin were analyzed in placentae of GDM. ResultsMorphological changes detected in GDM placentae included increased immature villi (16 cases, 38%), increased syncytial knots (36, 86%), perivillous fibrin deposition (20, 48%), fibrosis of villous stroma (20, 48%), presence of nucleated red blood cells (12, 28.5%) and hypervascularity (34, 81%). The extent of histopathological changes noted in GDM placentae was significantly higher than that in matched controls. GDM placentae showed significantly reduced expression of ANXA5 and Apelin in terms of grade and intensity when compared with matched controls. Reduced expression (mild intensity) of ANXA5 was noted in 22 GDM cases (52.3%) whereas apelin expression was of weak intensity in 26 (61.9%) cases. Among GDM patients, statistically significant association was noted between ANXA5 intensity and neonatal resuscitation, apelin grade and preterm birth as well as low birth weight and apelin intensity and requirement of treatment in sick neonatal care unit. ConclusionThe placental expression of the proteins, ANXA5 and Apelin, is altered in GDM though their exact pathogenetic mechanisms are yet to be understood. They can be targets for development of prophylactic and therapeutic agents in future.

Priming stroma with a vitamin D analog to optimize viroimmunotherapy for pancreatic cancer

Pancreatic cancer resistance to immunotherapies is partly due to deficits in tumor-infiltrating immune cells and stromal density. Combination therapies that modify stroma and recruit immune cells are needed. Vitamin D analogs such as calcipotriol (Cal) decrease fibrosis in pancreas stroma, thus allowing increased chemotherapy delivery. OVs infect, replicate in, and kill cancer cells and recruit immune cells to immunodeficient microenvironments. We investigated whether stromal modification with Cal would enhance oncolytic viroimmunotherapy using recombinant orthopoxvirus, CF33. We assessed effect of Cal on CF33 replication using pancreas ductal adenocarcinoma (PDAC) cell lines and in vivo flank orthotopic models. Proliferation assays showed that Cal did not alter viral replication. Less replication was seen in cell lines whose division was slowed by Cal, but this appeared proportional to cell proliferation. Three-dimensional in vitro models demonstrated decreased myofibroblast integrity after Cal treatment. Cal increased vascular lumen size and immune cell infiltration in subcutaneous models of PDAC and increased viral delivery and replication. Cal plus serial OV dosing in the syngeneic Pan02 model caused more significant tumor abrogation than other treatments. Cal-treated tumors had less dense fibrosis, enhanced immune cell infiltration, and decreased T cell exhaustion. Calcipotriol is a possible adjunct for CF33-based oncolytic viroimmunotherapy against PDAC.

Defective perlecan-associated basement membrane regeneration and altered modulation of transforming growth factor beta in corneal fibrosis.

In the cornea, the epithelial basement membrane (EBM) and corneal endothelial Descemet's basement membrane (DBM) critically regulate the localization, availability and, therefore, the functions of transforming growth factor (TGF)β1, TGFβ2, and platelet-derived growth factors (PDGF) that modulate myofibroblast development. Defective regeneration of the EBM, and notably diminished perlecan incorporation, occurs via several mechanisms and results in excessive and prolonged penetration of pro-fibrotic growth factors into the stroma. These growth factors drive mature myofibroblast development from both corneal fibroblasts and bone marrow-derived fibrocytes, and then the persistence of these myofibroblasts and the disordered collagens and other matrix materials they produce to generate stromal scarring fibrosis. Corneal stromal fibrosis often resolves completely if the inciting factor is removed and the BM regenerates. Similar defects in BM regeneration are likely associated with the development of fibrosis in other organs where perlecan has a critical role in the modulation of signaling by TGFβ1 and TGFβ2. Other BM components, such as collagen type IV and collagen type XIII, are also critical regulators of TGF beta (and other growth factors) in the cornea and other organs. After injury, BM components are dynamically secreted and assembled through the cooperation of neighboring cells-for example, the epithelial cells and keratocytes for the corneal EBM and corneal endothelial cells and keratocytes for the corneal DBM. One of the most critical functions of these reassembled BMs in all organs is to modulate the pro-fibrotic effects of TGFβs, PDGFs and other growth factors between tissues that comprise the organ.

Topographical distribution and histological characterization of stromal fibrosis in invasive breast carcinoma

Carcinoma breast is associated with various changes in organ and at the tissue level. The desmoplastic response in a tumor due to tumor hypoxia may result in dense or sparse, homogenous or focally hetrogenous fibrosis of the breast tissue. Peritumoral and stromal fibrosis has been assessed as an marker of intratumoral hypoxia and is being studied as histological surrogate prognostic parameter in breast cancer. We studied the topographical distribution and histological characterization of fibrosis and correlated it with various other histopathological parameters. 50 patients of carcinoma breast presenting to our institute were included in the study. We obtained a statistically significant correlation of presence of fibrosis with presence of lymphovascular invasion (p value =0.043), while we could not obtain any significant difference with age, menopause, tumor size, tumor grade, pathological stage, lymph node involvement, presence of necrosis, adipocytic invasion, duration of tumor, laterality, presence of inflammatory infiltrate and with ER, PR, HER2Neu status. This is a limited study in a small number of patients focussing on various stromal changes in breast cancer. This study has further characterized the heterogeneous nature of fibrosis in breast carcinoma using special staining which may facilitate the physician in making patient diagnosis, prognostication and outcome.

Clinical Images: Giant mass on the forehead in Kimura disease.

The patient, a 68-year-old Asian man with a history of nephrotic syndrome, presented to our department with multiple subcutaneous masses that developed over the past 50 years. The masses were painless, soft, and predominantly located in the head. The mass on the forehead was dome-shaped and had a diameter of 7 cm and a height of 3 cm (A). Blood tests showed eosinophilia and elevated immunoglobulin E (IgE) levels. Serum immunoglobulin G4 levels were normal. Ultrasonography showed heterogeneous masses with marked associated internal hyperemia (B). Magnetic resonance imaging showed T1-weighted isointense and T2-weighted hyperintense masses. A needle biopsy of the forehead mass showed stromal fibrosis, multiple lymphoid follicles, and infiltration of inflammatory cells, mainly eosinophils and lymphocytes. The patient was diagnosed with Kimura disease (KD). The masses shrank rapidly after administration of 30 mg/day of prednisolone for 2 weeks. KD is a rare chronic inflammatory disorder mainly affecting young Asian men. It presents as subcutaneous masses, typically in the head and neck, and is associated with eosinophilia and elevated IgE levels (1). It often affects the cervical lymph nodes and salivary glands (1). It is often associated with renal involvement, such as nephrotic syndrome (2). Glucocorticoids, surgical excision, and radiation therapy are the treatments of choice (3). The diagnosis of KD can be difficult because of its rarity. KD should be considered as a differential diagnosis when patients present with painless subcutaneous masses in the head and neck region with eosinophilia and elevated serum IgE levels.