Decrease In Spleen Size Research Articles

Liver Function Test Results Correlate With Spleen Size in Patients With Infectious Mononucleosis.

We aimed to evaluate the efficacy of measuring transaminase levels to determine the resolution of splenomegaly in athletes diagnosed with infectious mononucleosis (IM). Wecollected serial aspartate aminotransferase (AST) and alanine transaminase (ALT) levelsand ultrasound-measured spleen sizes in university athletes who had been diagnosed with IM. Our study included seven university-aged athletes from a single institution. Patients received serial liver function tests (LFT) and splenic ultrasound testing until resolution of symptoms and full return to sport. The effects of AST, ALT, and days from symptom onset were analyzed using multivariable mixed-effects linear regression models. Levels of AST and ALT were significantly correlated with spleen size. For each 10-unit increase in AST and ALT values, spleen size increased by 0.1 cm (p = 0.007) and 0.09 cm (p = 0.008), respectively. Decreasing levels of ALT and AST correlated with a decrease in spleen size. Normalization of AST/ALT values correlated with return of spleen size to baseline. Liver function testing may be useful in the return-to-play decision-making process for athletes with IM.

The Variation in the Traits Ameliorated by Inhibitors of JAK1/2, TGF-β, P-Selectin, and CXCR1/CXCR2 in the Gata1low Model Suggests That Myelofibrosis Should Be Treated by These Drugs in Combination.

Studies conducted on animal models have identified several therapeutic targets for myelofibrosis, the most severe of the myeloproliferative neoplasms. Unfortunately, many of the drugs which were effective in pre-clinical settings had modest efficacy when tested in the clinic. This discrepancy suggests that treatment for this disease requires combination therapies. To rationalize possible combinations, the efficacy in the Gata1low model of drugs currently used for these patients (the JAK1/2 inhibitor Ruxolitinib) was compared with that of drugs targeting other abnormalities, such as p27kip1 (Aplidin), TGF-β (SB431542, inhibiting ALK5 downstream to transforming growth factor beta (TGF-β) signaling and TGF-β trap AVID200), P-selectin (RB40.34), and CXCL1 (Reparixin, inhibiting the CXCL1 receptors CXCR1/2). The comparison was carried out by expressing the endpoints, which had either already been published or had been retrospectively obtained for this study, as the fold change of the values in the corresponding vehicles. In this model, only Ruxolitinib was found to decrease spleen size, only Aplidin and SB431542/AVID200 increased platelet counts, and with the exception of AVID200, all the inhibitors reduced fibrosis and microvessel density. The greatest effects were exerted by Reparixin, which also reduced TGF-β content. None of the drugs reduced osteopetrosis. These results suggest that future therapies for myelofibrosis should consider combining JAK1/2 inhibitors with drugs targeting hematopoietic stem cells (p27Kip1) or the pro-inflammatory milieu (TGF-β or CXCL1).

Reduction of Recipient Spleen Size after Living Donor Liver Transplantation: A Retrospective Study

Abstract Background Liver transplantation is considered an important solution to patients with end stage liver disease and a glimpse of hope for some patients with terrible quality of life due to the morbidity of such disease. Living donor liver transplantation procedures has now become widely done worldwide and many studies have been done to improve its outcome and improve the quality of the patients’ lives. Objective to help identify the factors that affects the size of spleen after living donor liver transplantation and promotes the reduction in its size which greatly affects the patients’ and graft survival rate. Patients and Methods This is a retrospective study including patients underwent living donor liver transplantation to be conducted in Ain Shams university hospitals to determine the factors promoting reduction of spleen size after living donor liver transplantation. This study was conducted at Ain-Shams University Hospitals, in the period from January 2008 to June 2021. Approval of the Ethical Committee and written informed consent from all participants were obtained. Results In our study we found that from 100 patients who had splenomegaly before transplantation, 54% of patients had persistent splenomegaly after liver transplantation, while 46% of patients had normal spleen size after transplantation within 1 year. The spleen size decreased significantly after transplantation in general, from a range of 13.3- 25 cm with a mean of 18.06 cm +/- 7 pre-op, to a range of 9-18 cm with a mean of 14.1 cm +/- 2.65. By analyzing this data and dividing it into two groups, it has been found that the group with persistent splenomegaly (54% of patients) had higher range of spleen size before transplantation with a range of 16.7-25 cm with a mean of 19.75 cm +/- 2.169, while the group of normal spleen size after transplantation ranged from 13.3-17 cm with a mean of 15.3 +/- 1.018 before transplantation. Also patients with dilated portal vein and portal vein thrombosis have higher chances of persistent splenomegaly post-operative. It has been found that in 93.4% of patients (43 patients)with reduced spleen size after transplantation had patent portal vein with average diameter, while only 6.5% (3 patients) had dilated portal vein and 0 patients with attenuated portal vein. On the other hand, 44.4% of patients with persistent splenomegaly had dilated portal vein, 38.9% had patent portal vein, and 16.67% had attenuated portal vein pre-op. Our study also showed the effect of spleen after transplantation on platelets levels and liver function. The normal spleen size was associated with better recovery of platelets number, in the group of patients with normal spleen size it ranged from 32 -189 10∧3/uL with a mean of 80 10∧3/uL pre-op, it increased to a mean of 163.95 10∧3/uL post op. While in the splenomegaly group the mean platelets count pre-op was 75.27 10∧3/uL in increased to 156.88 10∧3/uL post op. It has also been found that the decrease in spleen size greatly affects the liver functions, where in aspartate aminotransferase AST and Alanine transaminase ALT levels pre- operative in patients with normal spleen with a mean of 38.37 and 57.8 respectively. It decreased to a mean of 27 and 29.7 and difference of 11.2 and 28.1 respectively. Conclusion The spleen size after living donor liver transplantation is affected by the size of the spleen before transplantation and by the diameter portal vein before transplantation. Also the reduction in spleen size after living donor liver transplantation greatly affects the patient and graft survival platelets count and liver functions such as liver enzymes, albumin and coagulation profile.

MiR-146a-/- mice model reveals that NF-κB inhibition reverts inflammation-driven myelofibrosis-like phenotype.

Emerging evidence shows the crucial role of inflammation (particularly NF-κB pathway) in the development and progression of myelofibrosis (MF), becoming a promising therapeutic target. Furthermore, tailoring treatment with currently available JAK inhibitors (such as ruxolitinib or fedratinib) does not modify the natural history of the disease and has important limitations, including cytopenias. Since recent studies have highlighted the role of miR-146a, a negative regulator of the NF-κB pathway, in the pathogenesis of MF; here we used miR-146a-/- (KO) mice, a MF-like model lacking driver mutations, to investigate whether pharmacological inhibition of JAK/STAT and/or NF-κB pathways may reverse the myelofibrotic phenotype of these mice. Specifically, we tested the JAK1/2 inhibitor, ruxolitinib; the NF-κB inhibitor via IKKα/β, BMS-345541; both inhibitors in combination; or a dual inhibitor of both pathways (JAK2/IRAK1), pacritinib. Although all treatments decreased spleen size and partially recovered its architecture, only NF-κB inhibition, either using BMS-345541 (alone or in combination) or pacritinib, resulted in a reduction of extramedullary hematopoiesis, bone marrow (BM) fibrosis and osteosclerosis, along with an attenuation of the exacerbated inflammatory state (via IL-1β and TNFα). However, although dual inhibitor improved anemia and reversed thrombocytopenia, the combined therapy worsened anemia by inducing BM hypoplasia. Both therapeutic options reduced NF-κB and JAK/STAT signaling in a context of JAK2V617F-driven clonal hematopoiesis. Additionally, combined treatment reduced both COL1A1 and IL-6 production in an in vitro model mimicking JAK2-driven fibrosis. In conclusion, NF-κB inhibition reduces, in vitro and in vivo, disease burden and BM fibrosis, which could provide benefits in myelofibrosis patients.

Abstract 564: Therapeutic targeting of JAK/STAT signaling and histone acetyltransferase activity in post-myeloproliferative neoplasm secondary AML

Abstract Post-myeloproliferative neoplasm secondary Acute Myeloid Leukemia (Post-MPN sAML) is an aggressive and lethal hematologic malignancy arising from myeloproliferative neoplasms (MPN). Hyperactivation of JAK/STAT signaling was identified in 50-90% of MPNs - underscoring its role in driving MPN pathogenesis and transformation to post-MPN sAML. Ruxolitinib, a Type-I JAK2 inhibitor approved for the treatment of myelofibrosis, was tested in a Phase II clinical trial in post-MPN sAML. The trial revealed that ruxolitinib improved the quality of life in patients but was not disease-modifying. Thus, there is an urgent need to find synergistic drug combinations capable of complementing the therapeutic activity of JAK2 inhibitors in post-MPN sAML. Here, we demonstrate that combination treatment of ruxolitinib and CBP30, a bromodomain inhibitor of histone acetyltransferases (HAT) CBP and p300, enhances the therapeutic activity of ruxolitinib in post-MPN sAML. Genome-wide CRISPR loss-of-function screens in JAK2-mutated HEL cells determined that knockout of CBP sensitizes leukemic cells to JAK2 inhibitors. Consistently, functional validation through CRISPR/Cas9 demonstrated that genetic deletion of CBP but not of p300 renders HEL cells sensitive to ruxolitinib treatment. Pharmacologic inhibition of JAK/STAT signaling and HAT activity through ruxolitinib and CBP30 treatment in post-MPN sAML lines HEL and SET2 decreases proliferation and induces apoptosis and cell cycle arrest. Mechanistically, the synergism between ruxolitinib and CBP30 elicits a unique transcriptome profile in HEL with significant downregulation of E2F and MYC targets. In addition, epigenetic profiling revealed that the synergism induces global changes in H3K27ac signals with downregulation and upregulation of H3K27ac signals in predicted STAT5/BCL3 and GATA-bounded regions, respectively. Remarkably, combination treatment of ruxolitinib and CBP30 in HEL luciferase NSG mouse xenografts showed significant leukemia regression and decrease in spleen size compared to mice treated with vehicle or single agents. Overall, these results substantiate the potential of targeting JAK/STAT signaling and histone acetyltransferase activity in the treatment of post-MPN sAML. Citation Format: Kalay Bertulfo, Koen Debackere, Hannah Miller, Cindy Ma, Ryan Najac, Adolfo Ferrando, Teresa Palomero. Therapeutic targeting of JAK/STAT signaling and histone acetyltransferase activity in post-myeloproliferative neoplasm secondary AML [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 564.

CYSLTR1 antagonist inhibits Th17 cell differentiation by regulating the NF-κB signaling for the treatment of psoriasis.

Cysteinyl leukotriene receptor 1 (CYSLTR1) is observed to increase in psoriatic skin lesions. Montelukast, a CYSLTR1 antagonist, effectively treats inflammatory disorders, such as rheumatoid arthritis, multiple sclerosis, and atopic dermatitis. Thus, blocking CYSLTR1 may be a promising strategy for psoriasis immunotherapy. We prepared a montelukast sodium cream and solution and investigated their effects on psoriasis-like skin lesions induced by imiquimod (IMQ). After the treatment, serum, skin, and spleen samples were collected for evaluation. We treated human T helper (Th) 17 cells with montelukast in vitro to study its effect on Th17 differentiation and nuclear factor kappa-B (NF-κB) signaling. We also created a keratinocyte proliferation model induced by M5 cytokines and assessed the influence of montelukast on key psoriasis-related genes. We induced psoriasis in CYSLTR1 knockout (KO) mice using IMQ to explore the role of CYSLTR1 in psoriasis development. Montelukast sodium cream and solution effectively reduced the psoriasis area and severity index (PASI) and alleviated disease symptoms in IMQ-induced mice. Furthermore, reduced infiltration of inflammatory cells (Th1, Th17, and T follicular helper [Tfh] cells), decreased mRNA expression of cytokines in the skin (interleukin [IL]-17/F and IL-23), and lower serum concentrations of various cytokines (IL-2, IL-6, IL-13, and IL-17A/F) were observed. Montelukast cream and solution also decreased spleen size and the proportion of Th17 and Tfh cells, and significantly inhibited NF-κB signaling-related genes after application. Moreover, montelukast inhibited Th17 cell differentiation and suppressed NF-κB signaling in vitro. CYSLTR1 KO mice induced with IMQ showed improvement in PASI scores, serum IL-17A/F levels, and lower Th1 and Th17 cells in the spleen and skin compared to wild-type mice. Montelukast also suppressed the proliferation and inflammatory response of keratinocytes by regulating NF-κB signaling. Collectively, our results strongly indicate that inhibition of CYSLTR1 signaling to target the Th17 response holds significant promise as a therapeutic approach to manage psoriasis.

The mechanisms underlying alcohol-induced decreased splenic size: A network meta-analysis study.

Organ weight change is widely accepted as a measure of toxicologic pathology. We and other groups have shown that excessive alcohol exposure leads to decreased spleen weight in rodents. This study explores the mechanisms underlying alcohol-induced splenic injury through a network meta-analysis. QIAGEN Ingenuity Pathway Analysis (IPA) and Mammalian Phenotype (MP) Ontology were used to identify alcohol-related molecules associated with the small spleen phenotype. Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) and IPA bioinformatics tools were then used to analyze the biologic processes and enriched signaling pathways engaging these molecules. In addition, the "downstream effects analysis" algorithm was used to quantify alcohol's effects. IPA identified 623 molecules affected by alcohol and a Venn diagram revealed that 26 of these molecules overlapped with those associated with the MP Ontology of small spleen. The 26 molecules are TGFB1, CASP8, MTOR, ESR1, CXCR4, CAMK4, NFKBIA, DRD2, BCL2, FAS, PEBP1, TRAF2, ATM, IGHM, EDNRB, MDM2, GLRA1, PRF1, TLR7, IFNG, ALOX5, FOXO1, IL15, APOE, IKBKG, and RORA. Some of the 26 molecules were also associated with the MP Ontology of abnormal white pulp and red pulp morphology of the spleen, abnormal splenic cell ratio, decreased splenocyte number, abnormal spleen physiology, increased splenocyte apoptosis, and reduced splenocyte proliferation. STRING and IPA "Core Analysis" showed that these molecules were mainly involved in pathways related to cell apoptosis, proliferation, migration, and immune responses. IPA's "Molecular Activity Predictor" tool showed that concurrent effects of activation and inhibition of these molecules led to decreased spleen size by modulating apoptosis, proliferation, and migration of splenocytes. Our network meta-analysis revealed that excessive alcohol exposure can damage the spleen through a variety of molecular mechanisms, thereby affecting immune function and human health. We found that alcohol-mediated splenic atrophy is largely mediated by increased apoptosis signaling, migration of cells, and inhibition of splenocyte proliferation.

Jaktinib in patients (pts) with myelofibrosis (MF) who were refractory to or relapsed after ruxolitinib: A single-arm, open-label, multicenter, phase 2 study.

7062 Background: Currently, ruxolitinib is the only marketed Janus kinase inhibitor (JAKi) for pts with MF in China, which demonstrated symptomatic and splenic benefit with increased cytopenias. However, at least 50% of pts develop resistance or intolerance to ruxolitinib in the long-term follow up, for whom life expectancy is largely reduced. There are few therapeutic options after ruxolitinib failure. Jaktinib, an oral novel JAK and AVCR1 inhibitor showed promising activity in Jaki-naïve pts with intermediate or high risk MF with mild side effects. Here we present the results of jaktinib in MF pts who were refractory or relapsed to prior ruxolitinib therapy. Methods: Pts who had primary or post-ET/PV MF and were previously treated with ruxolitinib for at least 3 months and with either inadequate efficacy response (defined as < 10% spleen volume reduction [SVR] by MRI/CT or < 30% decrease in spleen size by palpation from baseline) or spleen regrowth to < 10% SVR or < 30% decrease in spleen size from baseline following an initial response were recruited to receive jaktinib 100 mg twice daily. The primary endpoint was the proportion of pts with SVR ≥ 35% from baseline (SVR35) at week (wk) 24, measured by MRI/CT images and assessed by Independent Review Committee. Secondary endpoints included the best spleen response rate (defined as achieving SVR35 at any time), the proportion of pts with ≥ 50% reduction in MPN Symptom Assessment Form Total Symptom Score (MPN-SAF TSS), improvement of anemia and safety. Results: A total of 34 pts were enrolled. The median duration of previous ruxolitinib treatment was 18.6 months (range 3.4-70.1). The median baseline TSS was 19.5 (range 1-55), hemoglobin (Hb) was 101.5 g/L (range 61-149), and platelets was 159×109/L (range 66-951). 11 pts (32.4%) achieved SVR35 at wk 24. The best spleen response rate was 38.2% (13/34). The median time to achieve the first SVR35 was 8.1 weeks, and the median duration of response was not reached. At wk 24, 13 pts (46.4%) achieved a ≥ 50% improvement in TSS from baseline. Of 16 transfusion-independent pts with Hb ≤ 100 g/L at baseline, 8 (50.0%) had a ≥20 g/L Hb increase by wk 24. Of 6 pts who required RBC transfusion at baseline, 2 had a 50% decrease in RBC infusion by wk 24. The median jaktinib exposure time was 231 days (range 96-371). 23 pts (67.6%) had one or more grade ≥3 treatment emergent adverse events (TEAEs). The most common grade ≥3 TEAEs were anemia (32.4%, n = 11), thrombocytopenia (32.4%, n = 11), leukocytosis (20.6%, n = 7) and leukopenia (11.8%, n = 4). Treatment discontinuation due to TEAEs occurred in 2 pts (5.9%). A dose reduction or temporary interruption due to TEAEs were reported in 9 pts (26.5%). Conclusions: MF pts who were refractory/relapsed to ruxolitinib attained substantial spleen response, and reduction in MF-related symptom burden with jaktinib, which could be a new option for MF pts who have failed ruxolitinib. Clinical trial information: NCT04851535 .

Dasatinib-induced spleen contraction leads to transient lymphocytosis.

The tyrosine kinase inhibitor dasatinib is approved for Philadelphia chromosome-positive leukemia, including chronic myeloid leukemia (CML). Although effective and well tolerated, patients typically exhibit a transient lymphocytosis after dasatinib uptake. To date, the underlying physiological process linking dasatinib to lymphocytosis remains unknown. Here, we used a small rodent model to examine the mechanism of dasatinib-induced lymphocytosis, focusing on lymphocyte trafficking into and out of secondary lymphoid organs. Our data indicate that lymphocyte homing to lymph nodes and spleen remained unaffected by dasatinib treatment. In contrast, dasatinib promoted lymphocyte egress from spleen with kinetics consistent with the observed lymphocytosis. Unexpectedly, dasatinib-induced lymphocyte egress occurred independently of canonical sphingosine-1-phosphate-mediated egress signals; instead, dasatinib treatment led to a decrease in spleen size, concomitant with increased splenic stromal cell contractility, as measured by myosin light chain phosphorylation. Accordingly, dasatinib-induced lymphocytosis was partially reversed by pharmacological inhibition of the contraction-promoting factor Rho-rho associated kinase. Finally, we uncovered a decrease in spleen size in patients with CML who showed lymphocytosis immediately after dasatinib treatment, and this reduction was proportional to the magnitude of lymphocytosis and dasatinib plasma levels. In summary, our work provides evidence that dasatinib-induced lymphocytosis is a consequence of drug-induced contractility of splenic stromal cells.

Spleen size change after hepatitis C treatment: a simple parameter to predict clinical outcomes.

Portal hypertension contributes to splenomegaly in cirrhotic patients. Reduction in spleen size may represent improvement in portal hypertension. The goal was to determine whether reduction in spleen size following sustained virologic response (SVR) in patients with hepatitis C virus (HCV) cirrhosis is associated with lower risk of liver-related adverse outcomes. A retrospective cohort study was performed regarding HCV-infected patients treated with direct-acting antiviral agents at the Iowa City Veterans Administration Medical Center between 2014 and 2019. Patients with cirrhosis and splenomegaly on baseline ultrasound were included. Spleen size, platelet counts, decompensations, hepatocellular carcinoma (HCC) status, and mortality were recorded through July 31, 2021. Decrease in spleen size ≥1.5 cm was regarded as significant. Intergroup comparisons were performed on SPSS 28. Eighty patients with cirrhosis and splenomegaly before SVR were identified. Spleen sizes decreased significantly after SVR in 31 patients over a median of 1 year (Group A), whereas 49 patients did not meet this endpoint (Group B). Lack of spleen size reduction was associated with the presence of varices before SVR (odds ratio (OR): 5.3, p < 0.01). Group A had significantly greater increases in platelet count after SVR than did Group B. Patients in Group B had greater risk of HCC (OR: 9.7, CI: 1.2-79; p = 0.03) and death (OR: 3.6, CI: 1.1-12; p = 0.04). Reduced spleen size in patients with HCV cirrhosis after SVR is associated with greater increment in platelet count, decreased risk of HCC, and reduced mortality compared to patients whose spleen size does not decrease.

Intraoperative portal vein stenting through umbilical vein approach: An innovative salvage procedure for portal vein thrombosis in pediatric liver transplant.

IPVS is considered a last resort or a salvage procedure in the event of recurrent PV thrombosis despite multiple attempts at redo PV anastomosis. We employed the opened umbilical vein approach to place the stent in the PV and deliver anticoagulation through a catheter. From Jan 2017 to Feb 2022, 150 patients underwent pediatric transplantation at department of liver transplant and hepatobiliary surgery unit, Indraprastha Apollo hospitals, New Delhi. Age, weight, PELD Score, diagnosis, portal vein diameter on preoperative CT, Portal flow after stenting, decrease in spleen size after stenting in follow-up CT were collected from a prospectively maintained data base and reviewed. Eight patients underwent IPVS following LDLT (mean age-10.6 ± 2.2 months, mean weight 8.1 ± 1.6, mean PELD score 32.7 ± 7.3). The mean PV diameter on preoperative CT scan was 3.6 mm (range 2.7-5.6 mm). The mean portal flow following stenting was 718.75 cc/min. Percentage reduction in size of the spleen was 26.35% beyond 2nd post-operative week. No patient had recurrent PV thrombosis following IPVS and all maintained an adequate portal flow throughout the immediate postoperative period. Two patients had in-hospital mortality secondary to septic complications. Umbilical vein approach is technically feasible, easy to manipulate the stent and catheter placement after stenting helps to deliver anticoagulants locally.

Pretransplant Splenic Irradiation in Patients With Myeloproliferative Neoplasms

PurposeAllogeneic hematopoietic cell transplantation (HCT) serves as the only curative treatment option for patients with myelofibrosis and other myeloproliferative neoplasms. Splenomegaly commonly manifests in patients with myeloproliferative neoplasms and can lead to delayed or poor engraftment, increased transfusion burden, and worse survival. Methods to decrease the effect of splenomegaly include splenectomy and splenic irradiation. We sought to report on clinical outcomes for patients treated with splenic irradiation as part of their transplant conditioning. Methods and MaterialsPatients with splenomegaly measuring greater than 22 cm were referred for splenic irradiation. They received radiation to the entire spleen to 10 Gy in 5 fractions using 3-dimensional conformal radiation with anteroposterior/posteroanterior or opposed tangent fields. Blood counts were monitored closely on treatment. Changes in splenic size were measured using first and last treatment image guided radiation therapy and pre- and posttransplant diagnostic imaging. ResultsSeventeen patients completed pretransplant splenic irradiation between 2012 and 2021. Median platelet, white blood cell, and hemoglobin levels decreased on treatment. One patient required platelet transfusion and 3 required packed red blood cell transfusions. Mean decrease in spleen size during radiation was -8.5% in the craniocaudal dimension. Prolonged decreases, measured 2 to 12 months after transplant, averaged 14.64%. All patients engrafted. Fourteen (82.4%) were alive at time of analysis with median follow-up of 4.2 years from hematopoietic cell transplantation. ConclusionsSplenic irradiation offers a safe method of managing significant splenomegaly as part of transplant conditioning. Transplant outcomes in this series were excellent. Prospective data may be beneficial to determine the absolute benefit of this addition to pretransplant conditioning in this patient population.

Cell erythroid maturation approach: a new paradigm in the road map towards a cure for β-thalassaemia syndromes.

Cell erythroid maturation approach: a new paradigm in the road map towards a cure for β-thalassaemia syndromes.

An Open-Label, Phase II Study Of The JAK1 Inhibitor INCB039110 In Patients With Myelofibrosis

An Open-Label, Phase II Study Of The JAK1 Inhibitor INCB039110 In Patients With Myelofibrosis

Evaluation of systemic involvement of Coronavirus disease 2019 through spleen; size and texture analysis

Background/aimTo investigate the changes in the spleen size, parenchymal heterogeneity, and computed tomography (CT) texture analysis features of patients diagnosed with Coronavirus disease 2019 (COVID-19)Materials and methodsThe size and parenchymal structure of the spleen in 91 patients who underwent thoracic CT examination due to COVID-19 were evaluated. For the evaluation of parenchymal heterogeneity, CT texture analysis was performed using dedicated software (Olea Medical, France). The texture analysis of each case consisted of 15 first-order intensity-based features, 17 gray level co-occurrence matrix-based features, and 9 gray level run length matrix-based features.ResultsA total of 91 patients (45 males, 46 females) with a mean age of 54.31 ± 16.33 years (range: 18–81) were included in the study. A statistically significant decrease in spleen size was seen in the follow-up CT examinations (p < 0.001) whereas no statistically significant difference was found between the Hounsfield unit (HU) values. The radiomics consisted of first-order intensity-based features such as 90th percentile, maximum, interquartile range, range, mean absolute deviation, standard deviation, and variance, all of which showed statistically significant differences (p - values: < 0.001, < 0.001, 0.001, 0.003, 0.001, 0.001, and 0.004, respectively). “Correlation” as a gray level co-occurrence matrix-based feature and “gray level nonuniformity” as a gray level run length matrix-based feature showed statistically differences (p-values: 0.033 and < 0.001, respectively).ConclusionsAlthough COVID-19 manifests with lung involvement in the early stage, it can also cause systemic involvement, and the spleen may be one of its target organs. A decrease in the spleen size and parenchymal microstructure changes can be observed in the short follow-up time. It is hoped that the changes in the parenchymal microstructure will be demonstrated by a noninvasive method: texture analysis.

Modulating Hedgehog Signaling Reduces MAPK, TGF-β, and Marrow Fibrosis in a Transgenic Mouse Model of JAK2V617F Associated Myeloproliferative Disease

Treatment of JAK2V617F driven myeloproliferative neoplasms (MPN) with FDA approved drug Ruxolitinib (JAK inhibitor, JAKi) has shown limited benefits. The failure of JAKi to eradicate JAK2V617F disease has led to the search for other pathways as potential therapeutic targets for this disease. Hedgehog (Hh) pathway inhibitors targeting Smoothened (SMO) have shown promising activity in MPN by reducing bone marrow fibrosis but are inadequate as single agents.In the present study we tested the effects of SMO inhibitor, PF-04449913 in transgenic mouse model of myeloproliferative disease (vavJak2V617F, gifted by Joe Zhao, University of Oklahoma Health Sciences Center). JAK2V617F transgenic mice exhibit a number of phenotypes that recapitulate those observed in MPN including bone marrow fibrosis and splenomegaly.Treatment with SMO inhibitor, PF-04449913, reduces splenomegaly. JAK2V617F mice showed pronounced splenomegaly compared to WT mice and treatment with SMO inhibitor resulted in a significant decrease in spleen size (Figure 1A). This was also associated with a drop in IL-4 and IL-7 in the peripheral blood (data not shown).SMO inhibitor reduces JAK2V617F allelic burden. In the next set of experiment we made mixed chimeras by transplanting one million JAK2V617F or WT bone marrow into lethally irradiated normal CD45.1 congenic hosts. Transplanted recipient mice were treated with PF-04449913 for one month and assessed for changes in peripheral blood chimerism. Treatment with PF-04449913 resulted in no change in WT engraftment. However, we found that PF-04449913 reduces the JAK2V617F allele burden in 3/5 mice.JAKV617F activates Hh signaling. Proteins were isolated from the spleens of 8-10 week JAK2V617F or WT mice. Western blot analyses demonstrate that JAK2V617F splenocytes express high SHH levels. We next measured levels of the GLI activator (GLI1) and the GLI repressor (GLI3R). Activation of Hh signaling is associated with an increase in GLI1 and a decrease in GLI3R. Treatment of the mice with PF-04449913 by daily gavage at 100mg/kg reduced GLI1 and increased GLI3R.JAK2V617F is associated with increased pERK, AKT, NF-κB activation. Oh and colleagues recently remonstrated the MPN were associated with activation of MAPK and NF-κB signaling pathways (Fisher et al., Leukemia, 2017). To understand how Hh signaling interacted with these pathways, we assessed ERK, AKT, and NF-κB activity by western blotting. JAK2V617F mice showed increased pERK, AKT and NFKB signaling. Treatment with PF-04449913 reduced pERK and AKT levels, confirming that both ERK and AKT are downstream of SMO, while it had little effect on IκBα levels suggesting that NF-κB activation occurs upstream of SMO.SMO inhibitor blocks bone marrow fibrosis by reducing TGF-β signaling. Numerous studies have implicated TGF-β in the pathogenesis of fibrosis. We analyzed the bone marrow of patients with JAK2V617F MPN and found them to express high TGF-β levels (Figure 1B). These findings were recapitulated in the JAK2V617F transgenic mice with levels of TGF-β in JAK2V617F splenocytes 15-fold greater than WT mice. Treatment with PF-04449913 normalized the levels of TGF-β (Figure 1C). Using a OP-9 co-culture system we demonstrated that JAK2V617F splenocytes robustly induced pSMAD2, which was completely blocked by PF-04449913 (Figure 1D). To assess for changes in bone marrow fibrosis, mice at 27 weeks were started on daily treatment with PF-04449913 for one month. Treatment with PF-04449913 blocks bone marrow fibrosis as demonstrated by decreased reticulin staining and reduced hydroxyproline. (Figure 1E-F).In conclusion, these data are the first to report that the Smoothened inhibitor, PF-04449913 modulates TGF-β and MAPK signaling in MPN and this results in decreased splenomegaly and marrow fibrosis. These results provide critical insight into the mechanism of action of SMO inhibitors in JAK2V617F associated MPN, and support the rational development of SMO and JAK inhibitor combinations in MPN.Figure Legend(A) SMO antagonist, PF-04449913 reduces splenomegaly. (B) Bone marrow of JAK2V617F MPN patient express high TGF-β levels. (C) Splenocytes of JAK2V617F transgenic mice express high TGF-β levels and PF-04449913 treatment normalizes TGF-β levels. (D) JAK2V617F induces Hh dependent TGF-β/pSMAD2 signaling in stroma. (E-F) PF-04449913 blocks fibrosis as measured by reticulin staining and quantification of Hydroxyproline. [Display omitted] DisclosuresO'Connell:Astex Pharmaceuticals, Inc.: Membership on an entity's Board of Directors or advisory committees; Otsuka: Membership on an entity's Board of Directors or advisory committees. Merchant:Pfizer: Consultancy, Research Funding.

RNAi and CRISPR/Cas9 Based In Vivo Models for Drug Discovery

With the advent of RNAi and more recently CRISPR/Cas9 technologies, we now have an advanced platform to rapidly generate disease and perform target validation and toxicity assessment of novel candidate targets all in the same animal. Having previously engineered a miRE scaffold for enhanced shRNA processing and now with the introduction of a powerful new SplashRNA algorithm for accurate shRNA prediction, we have brought RNAi technologies to its peak, by increasing potency and reducing off-target effects, such that it can be effectively exploited to preclinically mimic drug therapy and even combination therapy not only in vitro but also in live mice. Here, we take advantage of doxycycline-inducible RNAi mice to identify on-target toxicities associated inhibition of Rpl15 ,a novel gene target previously identified through screening as a potent therapeutic target for the treatment of HCC. We demonstrate that sustained Rpl15 suppression results in an impaired intestinal barrier with subsequent weight loss. Heterozygous mutation or deletion of Rpl15 has been associated with Diamond Blackfan Anemia in humans and we therefore analyzed our mice for hematological phenotypes. Peripheral blood counts revealed only a non-significant decrease in erythrocytes and hemoglobin in Rpl15 knockdown mice compared to mice expressing a control shRNA. However, we found that Rpl15 suppression resulted in a strong decrease in spleen size and histopathological analyses revealed signs of increased siderosis. Importantly, phenotypic toxicities are not observed following doxycycline removal and reexpression of Rpl15, thus suggesting that a therapeutic window of treatment exists for inhibition of this essential gene. By combining this approach with in situ delivery of Cas9/gRNAs, we proceed one step further to rapidly develop HCC and validate Rpl15 as potent target of HCC in vivo . These data showcase how synergizing our RNAi and CRISPR/Cas9 genetic toolbox will help facilitate drug discovery research and increase our confidence in predicting drug responses in humans into a new era. DisclosuresPremsrirut:Mirimus Inc.: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees. Wang:Mirimus Inc.: Employment. Yang:Mirimus Inc.: Employment. Fellmann:Mirimus Inc.: Consultancy, Equity Ownership. Dow:Mirimus Inc.: Consultancy. Zuber:Mirimus Inc.: Consultancy. Lowe:Mirimus Inc.: Consultancy.

Ultrasound criteria of splenomegaly in patients with fever and infectious mononucleosis

Ultrasound examination of the spleen is a promising approach allowing to accurately determine its normal size in various age groups as well as in pathological conditions. Objective – to evaluate spleen size changes in patients with febrile state and infectious mononucleosis. Ultrasound morphometric examination of the spleen was carried out in patients with febrile state (group “A”, n = 22) and infectious mononucleosis (group “B” n = 24) by assessing organ echo structure, contour clarity, and its uniformity. Morphometric data allowed to measure spleen weight and various coefficients by using the method proposed by Vozgoment et al. for calculating spleen weight (Wt) with formula: Wt = 0.34 * L2h, where L – the length of the spleen, h – the thickness of the spleen (cm); spleen weight coefficient (SWC) by using the formula: 1000 m / body weight (in grams), where m – the mass of the spleen; spleen weight-to-body height ratio (KI) by using the formula: spleen weight (g) / height (cm); spleen weight-to-body surface area ratio (Ks) by using the formula: spleen weight (g) / body surface area (m2). Statistical analysis was performed by using Statistica 10.0 software package. Febrile state in most cases is not accompanied by changes in spleen size. However, decreased spleen size was found in one (4.5%) patient aged 58 years, whereas splenomegaly – in 2 (9%) cases (SWC range 5.5- 5.9) in young patients aged 18 and 25 years. The course of infectious mononucleosis in 25% cases was not accompanied by change in spleen size. In other cases, both decreased (20.8%) and increased (54.2%) spleen size were observed. In particular, decreased spleen size was detected in patients with average age of 43.6±14.5 years, whereas increased spleen size (SWC range 4-8.9) was typical for young people aged 25±8 years. Febrile state in most cases is not paralleled altered spleen size. Infectious mononucleosis was associated with the three types of altered spleen size: no change, decreased spleen size characteristic to older patients, or increased spleen size observed in young subjects. Ultrasound examination of the spleen with measuring its morphometric parameters represent objective criteria allowing to fully assess the state of this immune organ and reveal degree of its involvement in the pathological process at the preclinical level.

MPN-340: Are There Any Age Differences in the Response to and Hematological Side Effects of Ruxolitinib Treatment in Myelofibrosis?

Objective: The JAK1/2 inhibitor ruxolitinib has been prescribed in myelofibrosis (MF) to reduce spleen size, but has been shown to be associated with drug-induced anemia and (or) thrombocytopenia. However, data regarding the use and safety profile of this drug in the elderly are scarce. Thus, our aim was to analyze the benefits and side effects of ruxolitinib in MF patients aged Methods: Cross-sectional, retrospective study conducted in the Department of Hematology, Center of Hematology and Bone Marrow Transplantation, Fundeni Clinical Institute, a tertiary reference hospital in Bucharest, Romania. Results: A total of 50 patients with MF were enrolled in our study: 50.0% (n=25) were aged Conclusions: According to our findings, elderly patients with MF frequently required a dose reduction to tolerate ruxolitinib treatment, whereas younger patients did not. Surprisingly, despite the dose reduction, the decrease in spleen size was more important in elder patients versus younger patients with MF. However, elderly MF patients also experienced a more significant reduction of the hemoglobin value while on ruxolitinib.

Effects of two little-studied environmental pollutants on early development in anurans

Despite intensive ecotoxicological research, we still know relatively little about the ecological impacts of many environmental contaminants. Filling these knowledge gaps is particularly important regarding amphibians, because they play significant roles in freshwater and terrestrial ecosystems, and their populations are declining worldwide. In this study, we investigated two pollutants that have been poorly studied in ecotoxicology despite their widespread occurrence in surface waters: the herbicide terbuthylazine and the pharmaceutical drug carbamazepine. We exposed two anuran species throughout their larval development to each of two environmentally relevant concentrations of each pollutant, and recorded mortality and 17 sub-lethal endpoints up to several months after exposure. Mortality was low and unrelated to treatment. In agile frogs (Rana dalmatina), we found that treatment with 0.3 μg/L terbuthylazine decreased tadpole activity and reduced fat bodies in juveniles, whereas treatment with 50 μg/L carbamazepine decreased spleen size and increased spleen pigmentation. In common toads (Bufo bufo), treatment with 0.003 μg/L terbuthylazine increased body mass at metamorphosis, treatment with 0.3 μg/L terbuthylazine increased the size of optic tecta, and treatment with 0.5 μg/L carbamazepine decreased hypothalamus size. Treatment with 50 μg/L carbamazepine reduced the feeding activity of toad tadpoles, decreased their production of anti-predatory bufadienolide toxins, and increased their body mass at metamorphosis; juvenile toads in this treatment group had reduced spleen pigmentation. Neither treatments affected the time to metamorphosis, post-metamorphic body mass, or sex ratios significantly. These results show that environmental levels of both terbuthylazine and carbamazepine can have several sub-lethal effects on anurans, which may be detrimental to individual fitness and population persistence in natural conditions. Our findings further highlight that toxic effects cannot be generalized between chemicals of similar structure, because the terbuthylazine effects we found do not conform with previously reported effects of atrazine, a related and extensively studied herbicide.