bluuebunny/crossref_metadata_2025_split huggingface dataset:
MicroRNA-92 expression in CD133+ melanoma stem cells regulates immunosuppression in the tumor microenvironment through integrin-dependent TGF-β activation
Authors: Shidal Chris , Nagarkatti Mitzi , Nagarkatti Prakash S. 
Published: January 2023

Abstract
The existence of cancer stem cells (CSC) accounts for the high degree of chemoresistance and heterogeneity characterizing refractory melanomas. Yet, the degree to which CSCs modulate immune cells in the tumor microenvironment has yet to be revealed. The present study aims to establish a novel role for miR-92 and its associated gene networks in immunomodulation. We employed syngeneic mouse models utilizing B16-F10 melanoma cells to observe primary tumor and metastatic growth. CSCs were isolated from the parental line based on expression of the putative CSC marker, CD133. CD133+ cells were functionally distinct from CD133− cells with CD133+ cells showing increased proliferation in vitro and enhanced tumorigenesis when administered subcutaneously. Our data indicated that compared to CD133− cells, CD133+ CSCs had a greater capacity to recruit immunosuppressive cell types during tumor formation. Using microarray technology in order to expose disparities in microRNA expression between CSC and non-CSC compartments, we identified several miRs that were significantly downregulated in CD133+ cells including miR-92. We hypothesized that lower levels of miR-92 in CSCs led to higher expression of integrin subunits as predicted by gene alignment software and confirmed using qRT-PCR. Flow cytometry analysis of dissociated tumors demonstrated that tumors initiated by CD133+ cells displayed significantly higher levels of TGF-β compared to CD133− cells. Additionally, qRT-PCR for TGF-β associated genes confirmed a significant increase in expression in CD133+ cells. These data suggest that CSCs efficiently induce intratumoral immunosuppression which is potentially regulated by a miR-92 driven axis involving integrin activation of TGF-β.
Targeting FAAH in hematological and non-hematological cancers
Authors: Doan Thao , Sun Jeff , Henderson Jared , Du Yong , Mohan Chandra 
Published: January 2023

Abstract

Background
FAAH is an enzyme that degrades endocannabanoids (eCBs), which have been shown to have pro-apoptotic effects on cancer cells. The body naturally produces eCBs, but they are degraded by endogenous FAAH. FAAH presence was investigated in diffuse large B-cell lymphoma (DLBCL) and breast cancer. By inhibiting the effects of FAAH through small molecule inhibitors, we hope to increase the effects of natural eCBs, such as AEA and PEA, in inducing apoptosis in cancer cells.

Methods
FAAH presence was measured through western blot in 8 DLBCL lines and 3 human breast cancer cell lines using anti-FAAH antibodies. Flow cytometry and MTT assays were used to measure apoptosis. Twenty nude mice were injected to 2M MDA-MB-231 GFP/luciferase cells in the mammary fat pad. After 7 days, tumors were imaged using IVIS and split into two groups. At 14 days, treatment with URB 597 (10mg/kg, 3 times a week) was commenced.

Results
Elevated FAAH was observed in DLBCL lines and breast cancer cell lines. Inhibition of FAAH and addition of exogenous eCBs resulted in decreased proliferation and increased necrosis of the tumor lines. The addition of exogenous eCBs and inhibition of FAAH resulted in a decrease in total EGFR, Akt, and Erk1/2 in MDA 231 tumor cells. There was also a decrease in p-Akt and p-Erk1/2. Inhibiting FAAH and eCB addition resulted in decreased cell viability (p &lt; 0.01) when compared to control with MDA 231 with AEA/URB. By tumor size (2.13 fold decrease, p &lt; 0.001), there was a reduction in in-vivo tumor growth over time with treatment using URB 597.

Conclusions
Inhibition of FAAH in vivo results in the buildup of eCBs, which activates apoptosis and results in decreased tumor growth. FAAH inhibition is a novel therapeutic approach for DLBCL and breast cancer.
Platelet activation alters the immune landscape of the tumor microenvironment
Authors: Riesenberg Brian P. , Rachidi Saleh , Metelli Alessandra , Gutierrez Jennifer , Liu Bie , Li Zihai 
Published: January 2023

Abstract
In a cancerous setting, the normal hemostatic role for platelets can be hijacked to promote tumor growth, survival and metastasis. The most well-established role for platelets in this regard occurs in the bloodstream where activated platelets coat and protect malignant tumor cells from immune attack allowing for successful metastases. Recently, platelet activation was shown to suppress the primary immune response to cancer, indicating a functional role for platelets in the tumor. However, the extent to which this phenomenon is generalizable or the degree to which platelets influence the immune signature of the tumor microenvironment has yet to be fully elucidated. We found that genetic inhibition of platelet activation was capable of slowing primary tumor growth in multiple murine tumor models. Prophylactic treatment with pharmacological platelet inhibitors, aspirin and clopidogrel, slowed tumor growth similar to the genetic model. Antiplatelet therapy mediated tumor control was dependent on the adaptive immune system, more specifically functional CD8+ T cells. Finally, we found therapeutic intervention targeting platelets in established tumors significantly altered the immune landscape of the microenvironment by increasing the percentage and altering the phenotype of CD8+ T cells. These data show that platelet activation can suppress the immune response to multiple cancers. Furthermore, this is the first study to demonstrate targeting platelet activation changes the immune landscape of tumor infiltrating lymphocytes suggesting a potential synergy with other immunotherapeutic strategies to improve tumor control.
Development of an Orthotopic Colorectal Cancer Mouse Model for Radiation Therapy Studies
Authors: Rodriguez Carlos J. , Battaglia Nicholas Griffin , Gerber Scott A. , Lord Edith M. 
Published: January 2023

Abstract
Radiation is commonly used before surgery to reduce tumor burden in rectal cancer. It has been suggested that radiation therapy induces an immune response against the tumor. Previous work using mice ectopic models has shown a correlation between tumor regression and the production of IFN-g and the induction of cytotoxic T cells after radiotherapy. Ectopic models, such as the use of colon cell lines in leg muscle, provide us with some information about the response to radiation, but this response may differ if the tumor is grown orthotopically in the tissue from which it originates. To assess this difference in response, we have developed an orthotopic model to study the growth of colorectal cancer in mice and the response to radiation treatment. In our model, MC38 adenocarcinoma murine cells expressing luciferase are implanted in the rectal wall of C57BL/6 mice through the serous membrane. Tumor growth, assessed using IVIS, revealed tumor masses that steadily increase in size reaching approximately 1 gram 21 days post-injection of 50,000 cells. Importantly, tumor growth in most mice was limited to the site of injection, as mesenteric and omental sites were free of tumor burden. The cellular composition and the morphology of these tumors is being examined using flow cytometry and immunohistochemistry, respectively. Furthermore, studies are underway to determine the response of these tumors to fractionated radiation and to determine if there are differences from the ectopic models of colon cancer. This more clinically relevant model will improve our understanding of response to radiotherapy in colorectal cancer and help develop more effective treatments for this disease.
Tumor cell-intrinsic programmed death protein 1 expression and induction in human cancer cell lines
Authors: Osta Erica , Gupta Harshita B. , Zhang Deyi , Kornepati Anand , Clark Curtis A. , Curiel Tyler J. 
Published: January 2023

Abstract
The programmed death protein 1 (PD-1) and ligand (PD-L1) axis inhibits T cell activation and reduces anti-tumor immunity. Most studies of the PD-1/PD-L1 axis test extrinsic, immune-dependent effects. However, tumor cell-intrinsic PD-L1 regulates immune-independent tumor mTOR signals, autophagy, and tumor initiating cell generation. Melanoma cell-intrinsic PD-1 regulates melanoma growth and mTOR. We tested PD-1 expression and induction in multiple human tumor cell lines. We detected PD-1 in human cell lines in vitro from ES2 ovarian cancer, VMM122 melanoma, and RT4 bladder cancer by flow cytometry, Western blot and RT-PCR. All lines were negative for the other PD-L1 receptor, CD80, by flow cytometry. Incubating lines with IFNα or IFNγ for 48 hours further augmented PD-1 expression as detected by flow, Western blot and RT-PCR. αPD-1 antibody (50 μg/ml) reduced proliferation of RT4 bladder cancer cells in vitro, demonstrating a direct PD-1 signal effect in these cells, and αPD-L1 similarly reduced in vitro proliferation. Interestingly, anti-PD-1 effects on reducing proliferation were similar: 41.5% reduction; p&lt;0.0001. We used CRISPR/Cas9 to generate PD-L1KO RT4 cells. αPD-L1 did not reduce in vitro proliferation as expected, but unexpectedly, αPD-1 also had no anti-proliferative effect on PD-L1KO RT4 cells, suggesting a PD-L1/PD-1 interaction. In murine B16 melanoma, we generated PD-L1KO and PD-1KO cells using CRISPR/Cas9. Cell-intrinsic PD-L1 and PD-1 downregulate chemokine production. Among chemokines analyzed, PD-L1 suppressed CCL2 and CCL5, PD-1 suppressed CCL4 and CCL7, and both suppressed CCL3 and CXCL2. Understanding intrinsic PD-1 and PD-L1 effects on tumor cells can be exploited to improve cancer immunotherapy
Focused ultrasound ablation as an immunomodulatory strategy for metastatic breast cancer therapy
Authors: Sheybani Natasha Diba , Witter Alexandra R. , Stevens Aaron D. , Bullock Timothy N. , Price Richard J. 
Published: January 2023

Abstract
Metastatic breast cancer (BrCa) is incurable, with a 5 year survival of only 22%. While immunotherapies are capable of generating durable responses in many forms of cancer, immune rejection of BrCa is very rare. Indeed, immunotherapies (e.g. checkpoint blockade) for BrCa are limited by poor functional CD8+ T cell infiltration and immunosuppression in the tumor microenvironment. Thus, adjunct strategies that render BrCa tumors responsive to immunotherapies are desperately needed. Focused ultrasound (FUS) mediates noninvasive acoustic energy deposition into tumors, conferring localized thermal and mechanical damage to targeted tissue. Here, we tested the hypothesis that FUS thermal ablation (TA) can serve as an auto-vaccine for treatment of BrCa with immunotherapy. Murine mammary carcinoma (4T1-HA) tumors were partially ablated with a 3 MHz ultrasound-guided FUS system. In tumor-draining lymph nodes, 1 week after FUS, we observed a ~3-fold increase in mature dendritic cells (CD11c-hi/CD86+), as well as an increasing trend in M1 macrophage representation by flow cytometry. While FUS did not confer changes in intratumoral CD8+ and CD4+ T cells, likely due to an overwhelming immunosuppressive burden imposed by myeloid derived suppressor cells (MDSCs), infiltrating T cells did express PD1. Therefore, we developed a combination treatment approach of FUS + gemcitabine and/or anti-PD1. This combination treatment elicited an ~50% reduction in 4T1-HA tumor growth at 5 days post-FUS ablation, suggesting that partial TA of BrCa tumors with FUS may represent a promising immunoadjuvant approach. Taken together, these results suggest immunoadjuvants abrogating immunosuppression may enable FUS to stimulate anti-tumor immunity in BrCa.
A delicate interplay between adaptive and innate immunity caused by immunotherapy triggers tumor immunity and aseptic inflammation
Authors: Hirschhorn Daniel , Ricca Jacob , Gasmi Billel , DeHanau Olivier , Mangarin Levi Mark , Budhu Sadna , Li Yanyn , Cortez Czarina , Liu Cailian , Zappasodi Roberta , Houghton Sean , Betof Allison , Lacouture Mario , Hollman Travis , Albrengues Jean , Egeblad Mikala , Wolchok Jedd D. , Merghoub Taha 
Published: January 2023

Abstract
Successful immune-based therapies that treat tumors are often accompanied by immune-related adverse events (irAE). Immunotherapy-treated patients who experience favorable anti-tumor responses typically manifest irAEs. The primary immunotherapies currently in clinic include agents that activate T cell responses such as checkpoint blockade of inhibitory pathways and infusion of ex-vivo tumor-derived or T cell receptor (TCR)-transgenic or chimeric antigen receptor (CAR)-modified T cells. While the beneficial and toxic effects of T cell-based immunotherapies in the clinic are being extensively explored, the precise mechanisms of tumor elimination and irAE remain the subject of intense investigation. In the present study, we treated established tumors with melanoma-specific adoptive CD4+ T cell transfer and costimulation via anti-OX40 or anti-CTLA-4. We found that, in spite of co-opting the adaptive immune response, acute local inflammation plays a fundamental role in tumor elimination and related toxicities in a model irAE. While stimulated T cells are necessary for initiating a therapeutic response, activation of endogenous neutrophils constitute an important and necessary effector mechanism of tumor destruction and irAE. Extensive neutrophils extracellular traps (NETs) were associated with irAE. Melanoma patients treated with checkpoint blockade, suffering from skin rashes equivalent to irEA found in mice, showed increased survival and NETois in biopsies from rashes and tumors. Our results bring forward a novel paradigm where T cells prime an anti-tumor immune response followed by an inflammatory effector mechanism, provided by the innate immune system with curative as well as morbid effects.
Differential abundance of mDC subsets predict response to Hepatitis B vaccination
Authors: Scheuermann Richard H. , Novotny Mark , Aevermann Brian , Ben-Othman Rym , Liu Aaron , Sadarangani Manish , Kollmann Tobias 
Published: January 2023

Abstract
Vaccination as a strategy for the prevention of infectious disease has been one of the greatest success stories in modern medicine, resulting in the complete eradication of smallpox, the near eradication of polio, and the drastic lowering in the incidences of measles, mumps, influenza and other common diseases. In contrast to these remarkable successes, recent failures in the development of effective vaccines against other major public health threats, including HIV, TB and malaria have highlighted the limitations to the current approach of empirical vaccine design and the need to better understand the basic principles of how to recognize and elicit immune responses that generate effective and durable protective immunity. Single cell transcriptional profiling by RNA sequencing (scRNAseq) is a powerful tool for exploring the phenotypic diversity of cell populations present in peripheral blood and other tissue specimens in an unbiased fashion. We applied scRNAseq to FACS sorted blood samples collected from participants before and after challenge with the licensed Hepatitis B vaccine, and identified several innate cell subtypes using unbiased clustering analysis. Marker genes specific for each cell subtype were identified using random forest machine learning for use in qPCR. Our results suggest that the relative levels of myeloid dendritic cells (mDCs) specifically expressing the N-myc target gene, NDRG2, prior to vaccination predict serum antibody responses to the HepB vaccine.
Intranasal immunization with an acellular pertussis vaccine containing a Th1/17 skewing adjuvant, BcfA, improves B. pertussis clearance from the mouse respiratory tract.
Authors: Yount Kacy S. , Jennings-Gee Jamie , Quataert Sally , Deora Rajendar , Dubey Purnima 
Published: January 2023

Abstract
Bordetella pertussis is the causative agent of whooping cough, a resurging vaccine-preventable disease. This highly contagious disease is most severe in infants and young children. Current alum-adjuvanted acellular pertussis vaccines (aPV) prevent severe disease but do not prevent nasal carriage and subsequent person-to-person pathogen transmission. The Th1/2 skewed immune response induced by aPV is one potential explanation for this failure. We are testing the hypothesis that modification of current aPV by the addition of an adjuvant, BcfA, that skews immune responses towards the more protective Th1/17 phenotype will improve protection. We also hypothesize that changing the delivery route from intramuscular (i.m) to intranasal (i.n) administration will generate mucosal immunity and further improve protection. In the established mouse model of B. pertussis infection, we found that i.m. immunization of mice with an experimental aPV containing BcfA and alum as the adjuvants significantly reduced bacterial numbers in the lungs compared to an alum-adjuvanted vaccine alone. Further, i.n administration of the BcfA+alum containing vaccine provided better protection against colonization of the trachea and lungs than i.m. immunization. Alum-induced Th2 cytokine responses were reduced by addition of BcfA, and were further reduced by i.n. vaccine delivery. Thus, a revised aPV that includes BcfA, administered i.n, may improve protection against B. pertussis infection compared with current formulations and delivery method.
Efficient Isolation of Monoclonal Antibodies from Antigen-specific Human Plasmablasts and Memory B Cells from a Pneumococcal Conjugate Vaccinee
Authors: Cox Kara S. , Chen Zhifeng , Tang Aimin , Fink Malorie , Guan Liming , Roman Jae , Verch Thorsten , Retzlaff Mary , Vora Kalpit A. 
Published: January 2023

Abstract
Community-acquired pneumonia is a leading infectious cause of hospitalization, the annual incidence of Pneumococcal pneumonia is 24.8 cases per 10,000 adults in USA. Pneumococcal conjugate vaccine (PCV13) is currently used in clinic to prevent pneumococcal disease. We isolated plasmablast cells from a Pneumococcal conjugate vaccinee 7 days post vaccination to understand humoral responses. The plasmablast cells were single-sorted into 96 well PCR plates and applied to direct molecular cloning for PCR amplification of antibody sequences. Paired antibody heavy and light chain variable region sequences were achieved from 35 out of 48 wells with molecular cloning success rate of 72.9%. The antibody sequences were recombinantly expressed as human IgG1, and 17 out of 35 hits were positive for vaccine antigen by ELSIA s giving a positive rate of 48.9%. We also isolated memory B cells from the same donor 28 days post vaccination. The memory B cells were single-sorted into 96 well cell culture plates pre-seeded with CD40L-expressing feeder cells in complete RPMI 1640 medium complemented with IL21 and cultured for two weeks. The culture supernatants were tested in ELISA and 7 wells were identified as antigen-specific hits. Paired heavy and light chain antibody sequences were amplified from all of the 7 hits, all of them were successfully cloned and recombinantly produced as human IgG1 and tested in ELISA. 6 out of the 7 hits (85.7%) turned out to be vaccine antigen specific. We will further compare and discuss the clonal specificities isolated from plasmablast and memory B cells to understand the clone’s participation in immune responses to vaccination.
Generating Neutralizing Antibody responses using Recombinant Infectious Rhinovirus
Authors: Lee Sujin , Tang Roderick , Currier Michael , Moore Martin 
Published: January 2023

Abstract
Human Rhinovirus (RV), a leading causative agent for common colds and the subsequent development of pneumonia, can exacerbate both asthma and chronic obstructive pulmonary disease (COPD). There is no vaccine against RV yet. As previous studies demonstrated that RV-neutralizing antibodies (nAb) correlates with protection, eliciting robust nAb responses might be a key factor for ideal RV vaccine development. Five neutralizing immunogenic (NIm) sites were identified. NIm-IA, IB, and IV are located on VP1, while NIm-II and III are located on VP2 and VP3, respectively. Since the capsid proteins (VP1, 2, 3 and 4) seem to be immunogenic, we hypothesize that a chimeric virus expressing the capsid proteins is sufficient to elicit nAb responses. The HRV-A76 insert expressing the four capsid proteins and 2A protein was cloned into a bacterial artificial chromosome (BAC) harboring cDNA expressing the full-length genome of HRV-A33. Utilizing reverse genetics, a capsid-chimeric recombinant RV strain, BAC HRV33cap76, was generated using the four capsid proteins and 2A protein from HRV-A76, and the remaining part of the genome from HRV-A33. Following RNA transfection and recovery, we found that the titer of BAC HRV33cap76 was similar to biological HRV33 or HRV76. We next immunized BALB/c mice intramuscularly with inactivated BAC HRV33cap76 mixed with alum adjuvant to determine its immunogenicity. The BAC HRV33cap76 immunization resulted in nAb against only HRV-A76. Our findings demonstrated that the capsid proteins are responsible for the generation of nAb against RV. As a capsid-chimeric recombinant RV was capable of eliciting robust nAb, it could be a novel vaccine platform to generate highly effective and safe vaccine for preventing RV infection.
Vaccination against Follicle Stimulating Hormone Receptor delays ovarian cancer progression by breaking immune tolerance
Authors: Puchalt Alfredo Perales , Wojtak Kryzstof , Duperret Elizabeth K. , Yan Jian , Weiner David B. 
Published: January 2023

Abstract
Ovarian cancer is a frequent disease with poor prognosis. Improvements in surgery and chemotherapy have not been able to improve the 5-year rate survival. The follicle stimulating hormone receptor (FSHR) is expressed in approximately 50% of ovarian cancers but not in healthy tissues except for the gonads. We generated a vaccine against FSHR consisting of a consensus sequence of FSHR from other species that has a 95% homology with murine FSHR. The FSHR vaccine was able to break immune tolerance generating a strong FSHR-specific CD8 and CD4 response. This response was long lasting and able to delay progression of FSHR+ tumors in an immunocompetent mouse. Vaccination generated an increased T-cell infiltration in the tumor microenvironment and was able to synergize with previously inactive anti-PD1 antibodies. We found that the newly generated FSHR-specific CD8 T cells were sufficient and necessary for the effect of our vaccine. In conclusion, vaccination against FSHR is a promising new immunotherapy against ovarian cancer and can increase the susceptibility of ovarian cancer to checkpoint blockade.