bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cigarette Smoking Exacerbates Flu-Induced Thrombo-Inflammation
Authors: Kaminski Tomasz W. , Brzoska Tomasz , Vats Ravi , Tutuncuoglu Egemen , Nickolich Kara , Robinson Keven M. , Nyunoya Toru , Sundd Prithu 
Published: November 2021

Abstract
Rationale: Epidemiological evidence suggests that prior exposure to cigarette smoke (CS) or habitual smoking increases the risk of influenza A virus (IAV)-triggered respiratory failure (severe flu). Although emerging evidence supports the role of thrombo-inflammation in the development of CS and IAV-triggered lung injury, the innate immune mechanism that contributes to this morbidity remains poorly understood.
Materials and methods: We have developed a two-hit model of CS-induced severe flu in mice. Mice were exposed to four weeks of room air (air) or CS followed by intra-nasal administration of A/PR/8/34 (H1N1) IAV. The body weight was measured every day for two weeks after IAV administration followed by assessment of lung injury at day 7 and 14. Lungs were harvested for histological assessment of lung injury and estimation of viral titer by RT-PCR. Quantitative fluorescence intravital lung microscopy (qFILM) was conducted at 2-, 3- and 4-days post IAV-infection to visualize dynamics of neutrophil and platelet recruitment in the lung of mice IV administered with fluorescent dextran, anti-Ly6G Ab and anti-CD49Ab.
Results: Mice exposed to CS+IAV manifested significantly more weight loss, lung injury, lung congestion and alveolar hemorrhage compared to mice administered room-air+IAV. QFILM revealed that severity of lung injury was associated with significantly more entrapment of neutrophil-platelet aggregates within the pulmonary microcirculation and infiltration into the air spaces of CS+IAV than room-air+IAV administered mice.
Conclusion: These initial results suggest that CS primes innate immune signaling in neutrophils and platelets to promote their recruitment in the lung following flu. Currently, studies are underway to identify innate immune pathways in neutrophils and platelets that drive this severe thrombo-inflammatory response.

Disclosures
Sundd: CSL Behring Inc: Research Funding; Bayer: Research Funding; Novartis: Membership on an entity's Board of Directors or advisory committees, Research Funding.
PD-L1 and Tumor Mutational Burden Are Not Predictive of Thrombosis in a Cohort of 1,221 Patients with Solid Organ Malignancies
Authors: Thawani Rajat , Kartika Thomas , Elstrott Benjamin , Batiuk Elizabeth , Chen Lilian , Tun Nattapron , Taflin Nicholas , Tao Derrick Law , West Malinda T , Shatzel Joseph J 
Published: November 2021

Abstract
Introduction: Malignancy is a well-known risk factor for thrombosis. While many clinical risk factors for cancer-associated thrombosis have been described, it remains unknown how certain tumor specific observations (such as programmed death ligand 1 expression (PD-L1) and tumor mutational burden (TMB)) correspond to thrombotic risk. To determine the relationship between PD-L1 and TMB quantification and thrombosis in patients with solid tumors, we evaluated a large cohort of clinical samples from an NCI designated cancer center, correlating tissue sample pathology with the development of venous or arterial thromboembolism.
Methods: We performed a retrospective cohort study of patients between the ages of 18-89 who underwent histopathologic examination and next generation sequencing for a diagnosis of a solid organ malignancy at the Knight Cancer Institute between June 2019 and Feb 2021. Medical records were reviewed to document clinical and demographic information as well as for the development of thrombosis after cancer diagnosis. Multivariable logistic regression was performed to assess if PD-L1 expression and TMB are independent predictors of thrombosis. Subgroup analysis was then performed to evaluate whether this relationship differed by the organ of the primary malignancy. All analyses were conducted in R (R Core Team 2019).
Results: We identified 1,221 patients with solid organ malignancies (mean age 62, 53.6% male). The most common malignancies in the cohort were lung cancer (13.8%), pancreatic cancer (12.3%) and colon cancer (12.3%). Thrombotic events occurred in 206 patients (16.8%) after diagnosis of their malignancy (100 deep vein thrombosis, 42 pulmonary embolism, 46 visceral vein thromboses, 6 superficial vein thromboses, and 11 arterial events). The mean time from initial biopsy to thrombosis was 224 days. TMB (mean 8.5 mutations/Mb) was evaluated in all patients in the cohort, while PD-L1 expression testing (mean 23.0%) was available for 255 patients. On multivariable logistic regression adjusting for age and patient sex, neither TMB (ORadj: 0.97, CI: 0.92-1.03) nor PD-L1 expression (ORadj 1.00, CI: 1.00-1.02) were significant predictors of thrombotic events. Subgroup analysis by primary malignancy type did not demonstrate any specific primary sites for which TMB or PD-L1 were predictive of thrombosis.
Discussion: Our analysis found no predictive relationship between TMB and PD-L1 expression with thrombotic events in patients with solid organ malignancy. Further analysis is needed to determine if specific treatment protocols, such as the use of immunotherapy in patients with varying levels of PD-L1 and TMB, alter thrombosis risk.

Disclosures
Shatzel: Aronora Inc,: Consultancy.
Fibrin(ogen) Mechanistically Contributes to Atopic Dermatitis Pathogenesis and Allergic Sensitization
Authors: Filuta Alyssa , Amezcua Peter K , Ruff Brandy , He Hua , Martin Lisa J , Palumbo Joseph S. , Khurana Hershey Gurjit K , Sherenian Michael G 
Published: November 2021

Abstract
Adults with atopic dermatitis (AD) have an increased risk for thromboembolic events. While coagulation is critical for almost all healing responses, dysregulated clotting and fibrin(ogen) deposition lead to inflammation that exacerbates tissue damage and impairs tissue repair. While plasma fibrin(ogen) has been associated with established asthma, fibrin(ogen)'s mechanistic role in AD pathogenesis and allergic sensitization has not been investigated. Here we show that fibrinogen plays a critical role in a murine model atopic dermatitis pathogenesis by markedly attenuating disease severity, barrier dysfunction, and allergic sensitization.
To determine if fibrinogen impacts AD pathogenesis, we used mice with complete fibrinogen deficiency (Fib -/-), partial deficiency (Fib +/-), and wildtype controls (Fib WT) in our established model of AD. This model uses repeated cutaneous allergen sensitization with heat-killed Aspergillus fumigatus (Asp) extract. Notably, Fib +/- mice produce approximately half as much fibrinogen as Fib WT mice. After each patch and at sacrifice each mouse underwent an objective disease severity assessment as well as transepidermal water loss (TEWL) measurements to assess skin barrier function. Our results show that complete and partial fibrinogen deficiency abrogated AD development. We found that Fib -/-and Fib +/- mice had markedly attenuated TEWL (P-value &lt;0.0001) and disease severity (P-value &lt;0.0001) compared with controls (Figure 1). In addition, we found that the TEWL and disease severity in Fib -/- and Fib +/- were comparable to unchallenged controls. There was also no difference in either outcome between Fib -/- and Fib +/- mice within the experimental group. Lastly, none of the Fib -/- and Fib +/- mice developed spontaneous bleeding.
Next, to elucidate the impact of fibrinogen on allergic sensitization we measured plasma allergen specific immunoglobulin E (sIgE) to Asp in all mice in our AD model at sacrifice. We found that Fib -/- and Fib +/- mice had significantly lower Asp sIgE compared with controls (p-value &lt;0.0001 and 0.0002, respectively; Figure 2). In addition, we found that the amount of sIgE to Asp produced was comparable between Fib -/- and Fib +/- mice as well as with unchallenged controls.
Thus, our findings suggest that fibrin(ogen) plays a critical mechanistic role in driving AD development and allergic sensitization. While fibrinogen's role in established allergic disease has been shown, these novel findings suggest a critical role for fibrinogen in allergic disease pathogenesis which may allow for additional therapeutic targets to treat atopic dermatitis and other allergic diseases.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Persistent Control of SIV Infection in Rhesus Macaques By Expressing a Highly Potent SIV Decoy Receptor after In Vivo HSC Transduction
Authors: Li Chang , Wang Hongjie , Gil Sucheol , Nelson Veronica , Kiem Hans-Peter , Lieber Andre 
Published: November 2021

Abstract
Despite the success achieved by antiretroviral HIV-1 therapies, long-term and repeated drug administration is associated with toxicity, virus evasion and high cost. We aim to develop a gene therapy approach for persistent control against HIV-1 infection by delivering the transgene expressing a decoy receptor (eCD4-Ig) to hematopoietic stem cells (HSCs) by in vivo transduction. In this approach, HSCs are mobilized from the bone marrow into the peripheral blood stream and transduced with intravenously injected virus vectors. We use an integrating, helper-dependent adenovirus (HDAd5/35++) vector system that targets human CD46, a receptor that is abundantly expressed on primitive HSCs. Transgene integration is achieved by a hyperactive Sleeping Beauty transposase and transgene marking in peripheral blood cells can be increased by in vivo selection. The efficacy and safety of our in vivo HSC transduction/selection strategy has been previously demonstrated for the treatment of b-hemoglobinopathies, hemophilia A, and cancer in murine disease models. In non-human primates, we showed efficient transgene (g-globin) expression in peripheral red blood cells using this strategy. eCD4-Ig functions like a neutralizing antibody and shows broad neutralization spectrum against HIV-1, HIV-2 and SIV isolates. We have designed and produced an HDAd5/35++ vector expressing rhesus eCD4-Ig (rh-eCD4-Ig) from a constitutive and highly active EF1a promoter. In CD46-transgenic mice, over 50µg/mL eCD4-Ig in serum was measured after in vivo transduction and selection, with no obvious adverse events observed. Neutralization assay with serum samples showed that the produced eCD4-Ig effectively inhibited HIV-1 and SIV infection. We then performed studies in a rhesus macaque. After in vivo HSC transduction/selection of a rhesus macaque, eCD4-Ig serum levels were stable at 20-30 mg/ml. In vitro SIVmac239 neutralization assays using week 13 serum and recombinant eCD4-Ig protein determined that the IC50 of eCD4-Ig in rhesus serum is 1.0 mg/ml. This implies that the serum eCD4-Ig concentration at the time of SIVmac239 challenge was ~25-fold higher than the IC50. High-level eCD4-Ig expression had no clinical or hematological side effects. The first SIVmac239 challenge (20pg) was given on June 22 nd. Increasing rechallenge doses are currently injected monthly. So far, the viral load measured by quantitative RT-PCR is below detection limit. The animal is without symptoms and has normal lymphocyte/subset counts. Our study demonstrates an in vivo HSC transduction approach for potential long-term control of HIV-1 infection.

Disclosures
Kiem: VOR Biopharma: Consultancy; Ensoma Inc.: Consultancy, Current holder of individual stocks in a privately-held company; Homology Medicines: Consultancy. Lieber: Ensoma: Research Funding.
Anti-Myeloma Effects of Panobinostat in Combination with Duvelisib
Authors: Chen Haiming , Li Mingjie , Goldwater Marissa , Behare Stacy , Cao Jasmin , Wallingford Cara , Hekmati Ava , Levin Rocky , Souther Eric , Xu Ning , Berenson James R. 
Published: November 2021

Abstract
Introduction: Panobinostat (Secura Bio) is a non-selective histone deacetylase inhibitor (HDACi) that has shown anti-tumor activity in preclinical studies in both solid and hematological malignancies. Duvelisib (Secura Bio) is an FDA-approved oral drug and a potent small molecule inhibitor of the delta (δ) and gamma (γ) isoforms of phosphoinositide-3 kinase (PI3K) with potential immunomodulating and antineoplastic activities. There is no data regarding in combination of an HDACi with duvelisib for multiple myeloma (MM) treatment. We evaluated the anti-myeloma effects of the panobinostat in combination with duvelisib.
Methods: The human MM cell lines U266, RPMI 8226 and MM1s were obtained from ATCC. Primary MM tumor cells were isolated from MM patients' bone marrow (BM) aspirates and mononuclear cells (MCs) were isolated. The cells were seeded at 10 5 cells/well in 96-well plates and incubated for 24 h in the presence of vehicle, panobinostat or duvelisib alone, or the two drugs together for 48 h. Cell viability was quantified using the MTS cell proliferation assay. For the apoptosis assay, U266, RPMI 8226, MM1s, or primary MM tumor cells were cultured at 1 x 10 6 cells per well for 48 hours at 37°C in 5% CO2 with or without the addition of panobinostat or duvelisib. Next, the cells were washed twice with PBS, re-suspended in binding buffer, and stained with FITC-conjugated annexin V and fluorescent dye propidium iodide (PrI) following annexin V assay standard protocol (BioVision, USA). For each drug treatment, 1 x 10 5 gated events were recorded.
Results and Discussion: The MTS cell proliferation assay showed that panobinostat alone inhibited cell proliferation of the MM cell lines U266 and MM1s but not RPMI8226. Panobinostat alone induced concentration-dependent inhibition of BMMCs. Panobinostat (20nM) in combination with duvelisib markedly increased inhibition of primary MM cell proliferation. Panobinostat alone induced MM cell apoptosis in all three MM cell lines. Panobinostat in combination with duvelisib enhanced MM cell apoptosis in MM1s but not U266 and RPMI 8226 cells. We further examined apoptosis of primary MM cell treated with panobinostat alone and in combination with duvelisib. The results showed panobinostat alone induced primary MM cell apoptosis and in combination with duvelisib markedly increased the induction of apoptosis. This study illustrates that the combination of panobinostat and duvelisib shows potent anti-MM effects in vitro and we are currently evaluating the anti-MM effects of this combination in vivo using our human MM xenograft models.
*The study was partially funded by Secura Bio, Inc

Disclosures
No relevant conflicts of interest to declare.
Development and Validation of a Novel Prognostic Nomogram Model and a Web-Based Survival Rate Calculator for Diffuse Large B-Cell Lymphoma in the Rituximab Era
Authors: Zhong Qiaofeng , Chen Haizhu , Qin Yan , He Xiaohui , Yang Sheng , Zhou Shengyu , Liu Peng , Shi Yuankai , Jing Hongmei 
Published: November 2021

Abstract
Objective This study aimed to develop a novel prognostic nomogram and a web-based survival rate calculator to improve discriminative and predictive accuracy for diffuse large B cell lymphoma (DLBCL) in rituximab era.
Methods We retrospectively collected data about 906 DLBCL patients who received R-CHOP or R-CHOP-like chemotherapy from January 2005 to December 2018 in National Cancer Center / Cancer Hospital, Chinese Academy of Medical Sciences. COX proportional hazard regression analysis was performed to assess the independent prognostic significance of clinical and histopathological variables on overall survival, which were used to develop the nomogram. We performed internal and external validation of the nomogram, and compared it's discriminative and predictive accuracy with the traditional IPI, R-IPI, and NCCN-IPI.
ResultsThe patients were divided into the training (n = 636) and validation (n = 270) cohorts at a ratio of 7:3. We identified six independent predictors for survival including age, ECOG, number of extranodal sites, bulky, and β2-MG, which were used to construct the nomogram and the web-based survival rate calculator. The C-index of the nomogram was 0.767 (95% CI, 0.735-0.799) in the training cohort and 0.732 (95% CI, 0.678-0.786) in the validation cohort. The AUC values of the nomogram for predicting the 1-, 5-, and 10-year OS rates for DLBCL were 0.794, 0.799, 0.805 in the training cohort and 0.828, 0.773, 0.771 in the validation cohort. All calibration curves revealed optimal consistency between predicted and actual survival both in the training and validation cohort. The nomogram divided DLBCL patients into three risk groups, the score of low risk, moderate risk, and high risk were 0-45，46-228，and higher than 228, respectively. The risk stratification model generated based on the nomogram showed a favorable level of predictive accuracy compared with the IPI, R-IPI, and NCCN-IPI in both cohorts according to the K-M survival curves. The AUC values of the nomogram model compared with the IPI, R-IPI, and NCCN-IPI for predicting the 10-year OS rates for DLBCL patients were 0.756 vs 0.748, 0.732, 0.713 in the training cohort and 0.742 vs 0.712, 0.713, 0.757 in the validation cohort.
ConclusionsIn conclusion, we have established and validated a novel prognostic nomogram model and a web-based survival rate calculator, which revealed more discriminative and predictive accuracy than IPI, R-IPI, and NCCN-IPI in the rituximab era.

Disclosures
No relevant conflicts of interest to declare.
ICG/LY@HES-PCL Nanomedicine Improving the Efficacy of CAR-T Therapy in Lymphoma
Authors: Tang Yuxiang , Mei Heng , Chen ZhaoZhao , Yao Weiqi , Li Chenggong , Xiong Wei , Hu Yu 
Published: November 2021

Abstract
Background :
The presence of tumor immunosuppressive microenvironment, abnormal vascular microenvironment, and the presence of a cross-linked matrix barrier may hinder the recruitment, proliferation, and activation of CAR-T to lymphoma tissue infiltration, which limit the efficacy of CAR-T in the treatment of lymphoma. The enhanced infiltration effect of ICG photothermal effect on CAR-T and the effect of LY on improving tumor immunosuppressive microenvironment have been confirmed. In this work, we have constructed LY/ICG@HES-PCL nanomedicine to improve the accumulation, proliferation and activation of CAR-T in lymphoma tissue, further improving the therapy efficacy of CAR-T in lymphoma.
Methods:
The effect of LY/ICG@HES-PCL on the proliferation and activation of CAR-T was studied by Transwell experiment. LY/ICG@HES-PCL was studied in vivo to investigate its improving the efficacy of CAR-T therapy in lymphoma. Biefly, 1×10 7 Raji human lymphoma cells were subcutaneously injected into both sides of each mouse. After ≈20 days, when the tumor volume reached ≈100 mm 3, mice were randomly assigned to 5 groups (n=6) and were treated by PBS, ICG@HES-PCL, ICG@HES-PCL+NIR, LY/ICG@HES-PCL, LY/ICG@HES-PCL+NIR, respectively. (4.0 mg/kg as ICG, 4.0 mg/kg as ICG). NIR groups were irradiated with an 808 nm laser at the power density of 0.3 W cm −2 for 10 mins. Two hours later, 1 × 10 7 CAR-T cells were intravenously injected into the mice. The tumor sizes were recorded by a digital caliper every 3 days. LY/ICG@HES-PCL in vivo was monitored using an in vivo bioluminescence imaging system. Lymphoma microenvironment was analyzed by immunofluorescence and FCM.
Results:
LY/ICG@HES-PCL increased CAR-T quantity by 3.7 times, IL-2 and IFN-λ release by 2.5 and 2.1 times in vitro, increased CAR-T accumulation in tumor tissue by 4.45 times within 3 days in vivo, achieving CAR-T activation, proliferation, and tumor accumulation enhancement. LY/ICG@HES-PCL improved CAR T efficacy in lymphoma by 3.4 times and CAR T longevity within 15 days by 1.6 times.
Discussion:
LY/ICG@HES-PCL co-delivered the photosensitifier ICG and TGF-β inhibitor LY to the lymphoma tissue. The photothermal effect of ICG decreased the density of lymphoma matrix and activated CD4+ T cells, CD8+ T cells and NK cells in tumor tissue. LY was used to inhibit PD-1 and CLTA-4 of CAR-T, so as to promotes CAR-T proliferation, improve CAR-T activity and longevity in vivo, and ultimately improve the anti-lymphoma efficacy of CAR-T cells.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
GSA: an independent development algorithm for calling copy number and detecting homologous recombination deficiency (HRD) from target capture sequencing
Authors: Chen Dongju , Shao Minghui , Meng Pei , Wang Chunli , Li Qi , Cai Yuhang , Song Chengcheng , Wang Xi , Shi Taiping 
Published: November 2021

Abstract
Background
The gain or loss of large chromosomal regions or even whole chromosomes is termed as genomic scarring and can be observed as copy number variations resulting from the failure of DNA damage repair.

Results
In this study, a new algorithm called genomic scar analysis (GSA) has developed and validated to calculate homologous recombination deficiency (HRD) score. The two critical submodules were tree recursion (TR) segmentation and filtering, and the estimation and correction of the tumor purity and ploidy. Then, this study evaluated the rationality of segmentation and genotype identification by the GSA algorithm and compared with other two algorithms, PureCN and ASCAT, found that the segmentation result of GSA algorithm was more logical. In addition, the results indicated that the GSA algorithm had an excellent predictive effect on tumor purity and ploidy, if the tumor purity was more than 20%. Furtherly, this study evaluated the HRD scores and BRCA1/2 deficiency status of 195 clinical samples, and the results indicated that the accuracy was 0.98 (comparing with Affymetrix OncoScan™ assay) and the sensitivity was 95.2% (comparing with BRCA1/2 deficiency status), both were well-behaved. Finally, HRD scores and 16 genes mutations (TP53 and 15 HRR pathway genes) were analyzed in 17 cell lines, the results showed that there was higher frequency in HRR pathway genes in high HRD score samples.

Conclusions
This new algorithm, named as GSA, could effectively and accurately calculate the purity and ploidy of tumor samples through NGS data, and then reflect the degree of genomic instability and large-scale copy number variations of tumor samples.
Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells
Authors: Rajabzadeh Alireza , Hamidieh Amir Ali , Rahbarizadeh Fatemeh 
Published: November 2021

Abstract
Background
Producing an appropriate number of engineered cells is considered as one of the influential factors in the successful treatments with chimeric antigen receptor (CAR) T cells. To this aim, the transduction rate of the viral vectors can play a significant role. In addition, improving transduction rates can affect the success rate of this treatment due to hard-transduced T lymphocytes.

Results
In this study, activated T cells were transduced using different transduction methods such as spinoculation, retronectin, polybrene, spinoculation + retronectin, and spinoculation + polybrene after selecting the most efficient transfection method to produce recombinant viral particles containing MUC1 CAR. PEI and lipofectamine with the amount of 73.72 and 72.53%, respectively, showed the highest transfection rates with respect to calcium phosphate (14.13%) for producing lentiviral particles. However, the cytotoxicity of transfection methods was not significantly different. Based on the results, spinoculation + retronectin leads to the highest transduction rates of T cells (63.19 ± 4.45%) relative to spinoculation + polybrene (34.6 ± 4.44%), polybrene (10.23 ± 0.79%), retronectin (10.37 ± 1.85%), and spinoculation (21.11 ± 1.55%). Further, the polybrene (40.02%) and spinoculation + polybrene (48.83% ± 4.83) increased cytotoxicity significantly compared to other groups.

Conclusion
Improving transduction conditions such as using spinoculation with retronectin can ameliorate the production of CAR-T cells by increasing the rate of transduction, as well as the success rate of treatment.