bluuebunny/crossref_metadata_2025 huggingface dataset:
Figure S9 from Antitumor Immune Responses in B2M-Deficient Cancers
Authors: Torrejon Davis Y. , Galvez Mildred , Abril-Rodriguez Gabriel , Campbell Katie M. , Medina Egmidio , Vega-Crespo Agustin , Kalbasi Anusha , Comin-Anduix Begoña , Ribas Antoni
Published: December 2023

B2M genetic alterations in pre-treatment human melanoma biopsies. Summary of B2M genetic alterations in pre-treatment human melanoma biopsies (n=295) determined by analyzing WES data processed by Sequenza. B2M homozygous loss n=2, LOH n=56, copy number gains n=88, unaltered n=149.
Data from A Metabolic Gene Survey Pinpoints Fucosylation as a Key Pathway Underlying the Suppressive Function of Regulatory T Cells in Cancer
Authors: Pinioti Sotiria , Sharma Himal , Flerin Nina C. , Yu Qian , Tzoumpa Amalia , Trusso Cafarello Sarah , De Bousser Elien , Callewaert Nico , Oldenhove Guillaume , Schlenner Susan , Thienpont Bernard , Garg Abhishek D. , Di Matteo Mario , Mazzone Massimiliano
Published: December 2023

Abstract
Forkhead box P3 (Foxp3)–expressing regulatory T cells (Treg) are the guardians of controlled immune reactions and prevent the development of autoimmune diseases. However, in the tumor context, their increased number suppresses antitumor immune responses, indicating the importance of understanding the mechanisms behind their function and stability. Metabolic reprogramming can affect Foxp3 regulation and, therefore, Treg suppressive function and fitness. Here, we performed a metabolic CRISPR/Cas9 screen and pinpointed novel candidate positive and negative metabolic regulators of Foxp3. Among the positive regulators, we revealed that targeting the GDP-fucose transporter Slc35c1, and more broadly fucosylation (Fuco), in Tregs compromises their proliferation and suppressive function both in vitro and in vivo, leading to alteration of the tumor microenvironment and impaired tumor progression and protumoral immune responses. Pharmacologic inhibition of Fuco dampened tumor immunosuppression mostly by targeting Tregs, thus resulting in reduced tumor growth. In order to substantiate these findings in humans, tumoral Tregs from patients with colorectal cancer were clustered on the basis of the expression of Fuco-related genes. FucoLOW Tregs were found to exhibit a more immunogenic profile compared with FucoHIGH Tregs. Furthermore, an enrichment of a FucoLOW signature, mainly derived from Tregs, correlated with better prognosis and response to immune checkpoint blockade in melanoma patients. In conclusion, Slc35c1-dependent Fuco is able to regulate the suppressive function of Tregs, and measuring its expression in Tregs might pave the way towards a useful biomarker model for patients with cancer.

See related Spotlight by Silveria and DuPage, p. 1570
Abstract A022: Immunotherapy of IL-6R Prevents Relapse and Metastasis of Triple-Negative Breast Cancer
Authors: Haq Aushia Tanzih Al , Yang Pao-Pao , Jin Christopher , Shih Jou-Ho , Chen Li-Mei , Wang Lu-Hai , Snyder Michael P. , Hsu Hsin-Ling
Published: December 2023

Multiple Copies in T-cell Malignancy-1 (MCT-1 or MCTS1) is a prognostic biomarker for aggressive breast cancers. Overexpressed MCT-1 stimulates the IL-6/IL-6R/gp130/STAT3 axis, promoting epithelial-to-mesenchymal transition, cancer cell stemness and tumor progression. To study the mechanism underlying TNBC immunity and aggressiveness, TNBC cell lines with different MCT-1 expression levels were evaluated. Primary tumor invasion and postsurgical local recurrence and distant metastasis were assessed in orthotopic syngeneic mice given indicated immunotherapy and/or MCT-1 silencing (shMCT-1). We found that shMCT-1 suppresses the transcriptome of inflammatory response and metastatic signaling in TNBC cells and inhibits TNBC progression, metastasis and mortality in xenograft mice. Combined IL-6R immunotherapy and shMCT-1 effect further decreased intratumoral M2 macrophages and T regulatory cells (Tregs) and avoided postsurgical TNBC expansion. Multiple positive feedback loops of the MCT-1/IL-6/IL-6R/CXCL7/PD-L1 axis were identified in TNBC cells, which boost metastatic niches and immunosuppressive microenvironments. MCT-1high/PD-L1high/CXCL7high and CXCL7high/IL-6high/IL-6Rhigh expression patterns predict worse prognosis and poorer survival in breast cancer patients. IL-6R-based immunotherapy more effectively prevented postsurgical TNBC metastasis, recurrence and mortality than anti-PD-L1 immunotherapy. Anti-IL-6R improved helper T, cytotoxic T and natural killer (NK) cells in lymphatic system and decreased Tregs in the recurrent and metastatic tumors, but anti-PD-L1 incapably elevated NK cells. Combined IL-6R and PD-L1 immunotherapies abridged TNBC cell stemness and M2 macrophage activity better than monotherapy. Sequential immunotherapy of PD-L1 and IL-6R demonstrated the best survival outcome, lowest postoperative recurrence and metastasis than synchronized therapy, particularly in shMCT-1 context. Conclusion: Systemic targeting of the MCT-1/IL-6/IL-6R/CXCL7/PD-L1 interconnection enhances the immune surveillance that inhibits aggressiveness of TNBC.

Citation Format: Aushia Tanzih Al Haq, Pao-Pao Yang, Christopher Jin, Jou-Ho Shih, Li-Mei Chen, Lu-Hai Wang, Michael P. Snyder, Hsin-Ling Hsu. Immunotherapy of IL-6R Prevents Relapse and Metastasis of Triple-Negative Breast Cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr A022.
Abstract A028: Defining immune pathways involved in the depletion of B-cell precursor acute lymphoblastic leukemia-initiating cells following early-life cytomegalovirus (CMV) infection in mice
Authors: Farrokhi Ali , Atre Tanmaya , Marquez1 Citlali , Gantt Soren , Reid Gregor
Published: December 2023

Background: B-cell acute lymphoblastic leukemia (B-ALL) is one of the most common forms of childhood cancers. Epidemiological studies have shown contrasting results regarding the role of infections in the risk of leukemia, and there is still no clear explanation for the pro- or anti-leukemia effects.

Aim: This study aimed to investigate the influence of cytomegalovirus (CMV) infection on the development of B-ALL and identify the underlying immune mechanisms involved.

Materials and Methods: In this study, Eμ-Ret transgenic mice were used as a model for hyperdiploid B-ALL, and luciferase-tagged CMV was used as a common early-life infection. The impact of infection on different subsets of B cells and leukemic initiating cells (LICs) in the bone marrow and spleen of neonatal and adult mice was assessed using multicolor flow cytometry. To gain insights into the mechanisms that impact LICs after infection, we sorted LICs from the splenocytes of infected and non-infected RFP mice seven days after infection, and total RNA sequencing (RNA-seq) was performed for gene expression analysis. We used neutralizing antibodies to evaluate the role of different cytokines/chemokines in mediating LIC depletion in vivo.

Results: Significant infection-induced depletion of preleukemic cells occurred in infected neonate Eμ-ret mice on BALBc, BALBc/FVB (F1) and BALBc/C57 (F1) backgrounds. This depletion showed an age-dependent pattern, with only infection in the first week after birth reducing the LICs numbers. Depletion of preleukemia was dependent on Stat4 and p40 homodimer/monomers, but not p40 heterodimers (IL-12 and IL-23). Several chemokines/cytokines, including IFNg and TNFa, showed high levels of expression in the serum of CMV-infected pups but not in adult mice. RNA sequencing indicated that IFNg and TNFa exerted a direct effect on preleukemic cells. In vivo neutralization of IFNg completely blocked the depletion effect of CMV infection. Further experiments confirmed that this cytokine works downstream of the Stat4 signaling pathway.

Conclusion: This is the first model to study the impact of CMV infection and the time of its exposure on B-ALL initiating cells. The identified Stat4-, IL12p40 homodimer/monomer-, and IFNg- driven activities could inform future treatment and prevention approaches for B-ALL.

Citation Format: Ali Farrokhi, Tanmaya Atre, Citlali Marquez1, Soren Gantt, Gregor Reid. Defining immune pathways involved in the depletion of B-cell precursor acute lymphoblastic leukemia-initiating cells following early-life cytomegalovirus (CMV) infection in mice [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr A028.
Abstract A031: Discovery and characterization of a novel, immunoproteasome activator that modulates the immunopeptidome, increases MHC class I antigenic presentation and enhances antitumor immunity
Authors: Driscoll James J. , Rana Priyanka S. , Ignatz-Hoover James J.
Published: December 2023

The transformative success of cancer immunotherapy has revolutionized cancer treatment and can induce long lasting responses in patients with cancers from a wide range of histologies. However, only a fraction of patients respond to immunotherapy and cancer cells escape immunosurveillance by downregulating antigen presentation through mechanisms that remain elusive. Proteasomes play a central role in the immune response by generating peptides from intracellular antigens which are presented on the cell surface for recognition by CD8+ cytotoxic T lymphocytes (CTLs). Immune cells contain a highly specialized variant of proteasomes, known as immunoproteasomes, in which the three constitutive catalytic subunits are replaced by cytokine-inducible, homologous catalytic subunits. Immunoproteasomes are specially adapted for a role in MHC class I antigen processing and presentation to CD8+ T-cells. Here, we performed a high-through screen (HTS) to detect novel molecular entities that activated proteasomal peptide-hydrolyzing activity. We identified a curated panel of molecules which not only enhanced proteasomal hydrolysis of the cell-permeable substrate LLVY-R110 but also enhanced the presentation of MHC class I antigens. E.G7-Ova is a mouse lymphoma cell line derived by electroporation of EL4 cells (C57BL/6, H-2b, T lymphoma) with chicken ovalbumin (OVA) cDNA. Proteasomes degrade OVA to generate the peptide SIINFEKL that is presented by MHC-class I molecules. Studies demonstrated that the hits identified in the screen increased SIINFEKL presentation on E.G7-Ova cells. The lead compound identified in the HTS, compound A, dramatically enhanced expression of the proteasome activator PA28, which is expressed by PSME1 and PSME2. Treatment of multiple myeloma (MM) cells with compound A enhanced the hydrolysis of Ac-ANW-MCA, which is selectively cleaved by the immunoproteasome beta5-associated catalytic subunits. Treatment of MM cells with compound A followed by mass spectrometry (MS) indicated that the relative presentation of individual antigenic peptides bound to MHC class I molecules on tumor cells was dramatically increased. Novel immunoproteasome-selective therapeutics have the potential to generate neoantigens and upregulate cspecific antigens on tumor cells to enhance CD8+ T-cell-mediated control of human cancers.

Citation Format: James J. Driscoll, Priyanka S. Rana, James J. Ignatz-Hoover. Discovery and characterization of a novel, immunoproteasome activator that modulates the immunopeptidome, increases MHC class I antigenic presentation and enhances antitumor immunity [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr A031.
Abstract A038: Targeting TCTP overcomes resistance to T cell-mediated immunotherapy by reversing the multi-malignant phenotypes of immune-refractory tumor cells
Authors: Kim Tae Woo
Published: December 2023

Cancer immunotherapy, particularly T cell-mediated therapy such as immune checkpoint blockade (ICB) and adoptive T cell transfer (ACT), has shifted the paradigm for cancer treatment. However, the emergence of immune-refractory tumor has limited its clinical success by disrupting one or more steps of the cancer immunity cycle. In addition, multi-malignant phenotypes of immune-refractory tumor cells are also one of the major causes result poor prognosis of patients. We need to identify novel predictive biomarker allowing for the selection of patients who will receive clinical benefit from immune-refractory tumors. Therefore, identifying the immune-resistance and multi-malignant factor, which not only can be targeted by clinically available medicines is very important. Here, we identified TCTP as a novel factor conferring multi-malignant phenotypes of immune-refractory tumor cells. We discovered a vital role of TCTP at the crossroads between multi-malignant tumor cells and the anti-cancer immunity system by demonstrating that TCTPhigh tumor cells enriched by immune selection pressure drive immune-refractory phenotypes. Importantly, the expression levels of TCTP within the tumors significantly associated with the clinical response of anti-PD-L1 therapy, which suggest TCTP as a prognostic marker in the case of clinical trials. Furthermore, targeting TCTP by clinical available drug overcomes the resistance to T cell-mediated therapy including ICB and ACT. Thus, our findings demonstrated that TCTP could be a both a valid target a prognostic marker providing a framework for patient selection to apply combined therapy of T cell-mediated therapy with TCTP-targeting drugs.

Citation Format: Tae Woo Kim. Targeting TCTP overcomes resistance to T cell-mediated immunotherapy by reversing the multi-malignant phenotypes of immune-refractory tumor cells [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr A038.
Abstract A054: Anti-CTLA-4 generates memory T-cell with greater expansion and functionality than anti-PD-1
Authors: Mok Stephen , Liu Huey , Anang Nana-Ama A.S. , Mancuso James J , Çobanoğlu Didem Ağaç , Allison James P
Published: December 2023

Blocking either cytotoxic T-lymphocyte antigen-4 (CTLA-4) or programmed cell death-1 (PD-1) pathway results in distinct T-cell differentiation. We investigated the effects of anti-CTLA-4 and anti-PD-1 on memory T-cell formation in mice. Anti-CTLA-4 generates a more potent memory response than anti-PD-1. Memory T-cells generated by anti-CTLA-4 expand at a greater frequency, have greater cytokine production, and exhibit higher antitumor activity than those generated by anti-PD-1. These memory cells also more frequently differentiate into KLRG1+ effector CD8 T-cells during re-challenge. Additionally, anti-CTLA-4 generates more TCF-1+ memory-like T-cells, while anti-PD-1 results in more TOX+ terminally differentiated T-cells. Together, we provide insights into the long-term effects of these immunotherapies on the immune system.

Citation Format: Stephen Mok, Huey Liu, Nana-Ama A.S. Anang, James J Mancuso, Didem Ağaç Çobanoğlu, James P Allison. Anti-CTLA-4 generates memory T-cell with greater expansion and functionality than anti-PD-1 [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr A054.
Abstract B002: Vidutolimod, an immunostimulatory virus-like particle, reduces proliferation but enhances the activation of tumor-specific T cells
Authors: Fischer Travis D. , Lemke-Miltner Caitlin , Weiner George J.
Published: December 2023

One approach to enhancing the anti-tumor T cell response is to alter the tumor microenvironment (TME) through intratumoral injection (IT) of immunostimulatory agents such as Vidutolimod (Vidu). Vidu is a virus-like particle (VLP) composed of a TLR9 agonist (CpG-A, known as G10) encapsulated by the Qβ bacteriophage capsid. IT Vidu shows considerable promise in early phase clinical trials. The immune response to Vidu is initiated by induction of IFNa production by pDCs within the TME. This effect is dependent on coating of Vidu with antibodies against the Qb capsid. This is followed by a series of changes in the TME that ultimately result in an enhanced anti-tumor T cell response. Mouse models have shown that the efficacy of IT Vidu depends on the presence of both CD4+ and CD8+ T cells. Most studies to date exploring the impact of Vidu on T cells have focused on the overall T cell population. The current studies were designed to further assess the complex mechanisms by which Vidu induces an anti-tumor T cell response through use of the well-established OT-1 mouse model that allows for analysis of the tumor-specific T cell population. OT-1 mice contain CD8+ T cells with a transgenic TCR that recognizes the ovalbumin (OVA) peptide SIINFEKL sequence (OVA257-264) presented on MHC Class I. Prior to culture, OT-1 splenocytes were labeled with CellTrace Violet in order to monitor proliferation over time. OT-1 splenocytes were then cultured with EL4 cells (an OVA-negative T lymphoblast cell line) or E.G7-OVA (OVA-expressing EL4 derivative cells). Minimal proliferation or evidence of T cell activation was seen when OT-1 CD8+ T cells were cultured with EL4 cells regardless of the addition of Vidu and anti-Qβ antibodies. OT-1 CD8+ T cells cultured with E.G7-OVA cells showed both proliferation and activation as indicated by increased intracellular IFNy and surface PD-1. Addition of Vidu and anti-Qb antibody reduced OT-1 CD8+ proliferation but enhanced production of IFNγ and expression of PD-1. The increase in IFNγ and PD-1 expression was strongest in the dividing OT-1 CD8+ T cell population. Preliminary results of ongoing in vivo studies are consistent with these results. In summary, Vidu reduces proliferation but enhances phenotypic markers of activation expressed by tumor-specific CD8+ T cells (OT-1 cells) when co-cultured with cells expressing OVA, their target antigen. Markers of activation are most notable in dividing OT-1 CD8+ T cells.

Citation Format: Travis D. Fischer, Caitlin Lemke-Miltner, George J. Weiner. Vidutolimod, an immunostimulatory virus-like particle, reduces proliferation but enhances the activation of tumor-specific T cells [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr B002.
Abstract B021: In vivo gain-of-function screen identifies CREB5, a novel ECM modulator that promotes immunotherapy resistance via the Collagen-Lair1 axis
Authors: Tiwari Payal , Colvin Kayla , Kim Sarah , Kammula Ashwin , Anderson Seth , Revach Or-Yam , Vyas Maulik , Talamas Jessica A , Demehri Shadmehr , Yates Kathleen , Doench John , Hahn William C , Manguso Robert
Published: December 2023

Treatment with immune checkpoint inhibitors induce remarkable clinical responses in several cancer types. However, most cancer patients do not respond to immunotherapy, and patients who initially respond often exhibit acquired resistance. Understanding the universe of immune evasion strategies will enable the design of more effective immunotherapies. Here, we identified genes that drive immune evasion by performing a genome scale in vivo CRISPR gain-of-function screen in tumors treated with anti-PD-1 antibodies; and discovered the transcription factor CREB5 as a top resistance mediator. Using transcriptional profiling, we showed CREB5 drives upregulation of extracellular matrix genes including collagen and collagen-stabilizing factors. We found CREB5 or collagen (Col1a1, Col4a1, or Col16a1) overexpressing tumors exhibit poor responses to anti-PD-1. Collagen is the major ligand for the inhibitory receptor LAIR1, which is broadly expressed on T cells, B cells, NK cells, and myeloid cells. Deletion of LAIR1 in mice or overexpression of the decoy receptor LAIR2 in tumors abrogates the resistance-causing effect of CREB5 or collagen overexpression, suggesting that CREB5 overexpression drives resistance partly via collagen-LAIR1 inhibitory signaling. In summary, we have identified a gene that can induce fibroblast-like features in tumor cells to promote immunotherapy resistance. Further investigation of pathways that induces stromal mimicry in cancer cells could identify novel immunotherapy targets.

Citation Format: Payal Tiwari, Kayla Colvin, Sarah Kim, Ashwin Kammula, Seth Anderson, Or-Yam Revach, Maulik Vyas, Jessica A Talamas, Shadmehr Demehri, Kathleen Yates, John Doench, William C Hahn, Robert Manguso. In vivo gain-of-function screen identifies CREB5, a novel ECM modulator that promotes immunotherapy resistance via the Collagen-Lair1 axis [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr B021.
Abstract B037: Single-cell analysis reveals immune characteristics linked to poor prognosis in Fusobacterium nucleatum-infected colorectal cancer
Authors: Choi Ilseok , Kim Kyung-A , Han Yoon Dae , Kim Sang Cheol , Kim Han Sang , Lee Insuk
Published: December 2023

Fusobacterium nucleatum (Fn) is commonly detected in colorectal cancer (CRC) and worsens patient survival. Fn appears to play a role in colorectal cancer carcinogenesis through suppression of the antitumor immune response. Aiming to uncover the underlying mechanisms, we collected 42 samples of surgically removed colon tissues from patients newly diagnosed with colon cancer. To analyze the bacterial community composition within these tissues, we utilized amplicon sequencing, targeting the V4 variable region of the 16S rRNA. We performed single-cell RNA sequencing (scRNA-seq) analysis of tumor-infiltrating immune cells from Fn-infected [Fn (+)] and Fn-uninfected [Fn (−)] patients. By utilizing gene expression signature derived from scRNA-seq data and bulk transcriptome profiles of the TCGA cohort, we identified immune cell types associated with Fn infection in CRC. Using RNA velocity and cell-cell interaction analysis of single-cell transcriptome data, we unraveled immune cell subtypes modulated by Fn infection. Furthermore, trajectory-based differential expression analysis and single-cell gene network analysis shed light on how the intratumor immune system is disturbed by Fn infection. A novel gene signature had a poor prognostic impact in patients with Fn-infected tumors, compared with Fn-uninfected patients in the TCGA cohort. Overall, this study identified a novel potential Fn-related immune evasion mechanism, beyond suppression of T cell-mediated immune response. Novel gene signatures with a poor prognostic impact can be used for patient stratification and developing targeted strategies in patients with Fn infection.

Citation Format: Ilseok Choi, Kyung-A Kim, Yoon Dae Han, Sang Cheol Kim, Han Sang Kim, Insuk Lee. Single-cell analysis reveals immune characteristics linked to poor prognosis in Fusobacterium nucleatum-infected colorectal cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr B037.
Abstract B045: EV-mediated immune modulation of CD8+ T cells in renal cell carcinoma
Authors: Jaschkowitz Greta , Noessner Elfriede , Zaccagnino Angela , Stöckle Michael , Junker Kerstin , Zeuschner Philip
Published: December 2023

Introduction: Treatment regimens for advanced renal cell carcinoma (RCC) involve immune checkpoint inhibition targeting the PD-(L)1 pathway. PD-L1 is often overexpressed on tumor cells in advanced disease and plays a crucial role in regulating tumor-directed immune responses. It can also be expressed on tumor-secreted extracellular vesicles (EVs), which are an essential part of intercellular communication. PD-L1 carrying EVs have been shown to have immunomodulatory effects in several tumor entities, however their role in RCC is still unknown. The aim of this study was the evaluation of putative immunomodulatory effects of RCC-derived small EVs on CD8+ T cells.

Materials and Methods: EVs of 5 untreated as well as interferon γ (IFNγ)-treated RCC cell lines were isolated through serial ultracentrifugation. Quality and quantity of the EVs were verified using Western Blotting (WB), Nano Particle Tracking Analysis and Transmission Electron microscopy, their PD-L1 expression was analyzed semi quantitatively through WB. CD8+ T cells were isolated from the blood of healthy donors, activated and subsequently co-cultured with RCC-derived EVs for a duration of 3 days. Proliferation and PD-1 expression of the T cells was assessed using flow cytometry. ELISAs were performed to analyze cyto- and chemokine secretion.

Results: All RCC cell lines expressed PD-L1, albeit at varying low levels. Four out of 5 treated cell lines responded to IFNγ treatment with an upregulation of PD-L1 expression on both cells and secreted EVs. Co-cultures of CD8+ T cells and RCC-derived EVs, particularly PD-L1+ EVs isolated from IFNγ-stimulated cells, lead to a reduction in T cell PD-1 expression. Additionally, PD-L1-expressing EVs of RCC cell line RCC53 had an antiproliferative effect on CD8+ T cells. EVs also increased the secretion of IL-2, IL-10, CXCL10 and CCL5 by the CD8+ T cells.

Conclusions: PD-L1 is expressed at low levels in RCC cell lines. However, repetitive IFNγ stimulation can lead to upregulation of PD-L1 on both cells and EVs enabling us to use this model to further investigate EV-mediated, PD-L1 dependent effects. In co-cultures, RCC-derived EVs impede early T cell activation and proliferation patterns of CD8+ T cells. Furthermore, early results indicate a shift towards a more pro-inflammatory cyto- and chemokine production by the T cells induced by RCC-derived EVs. The results support the hypothesis of a tumor-EV-mediated immune modulation in RCC.

Citation Format: Greta Jaschkowitz, Elfriede Noessner, Angela Zaccagnino, Michael Stöckle, Kerstin Junker, Philip Zeuschner. EV-mediated immune modulation of CD8+ T cells in renal cell carcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr B045.
Abstract B049: Lipid nanoparticle-delivered toll-like receptor agonists are highly effective in cancer immunotherapy
Authors: Bai Jun , Su Fei , Li Chen , Wu Yujing , Li Jing , Zhao Xiaobin , Yang Yongsheng , Lee Robert J
Published: December 2023

Background: Tumor immunotherapy has shown promising efficacy in various types of cancer. However, the effectiveness of immunotherapy is often limited by the immune-suppressive microenvironment within tumors. To address this challenge, we developed a method of intratumoral injection using lipid nanoparticles (LNPs) to deliver Toll-like receptor 7, 8, and 9 agonists. LNPs effectively enhance immune activation while reducing systemic responses by increasing drug retention within the tumor.

Methods: We encapsulated 30nt-CpG and Resiquimod into LNPs (QTsomeTM). Subcutaneous inoculation of 2 × 106 MC38 cells was performed in mice. Once the average tumor volume reached approximately 100mm3, we divided 15 mice into 3 groups based on tumor volume. Intratumoral injections began on the same day of grouping, with a dosage of 1mg/kg (5 injections every 3 days). Tumor volume and body weight were measured twice a week. We conducted two parallel experiments, each with 7 mice for the MC38 model and 8 mice for the B16 melanoma model in each group, to compare the efficacy of 26nt-CpG and 30nt-CpG.

Results: On the 15th day post-dosing, the tumor growth inhibition rate (TGI) was 79.9% (P<0.001), resulting in a 73.7% increase in survival rate. LNP-delivered 26nt-CpG demonstrated a TGI rate of 75.4% (P<0.001), while 30nt-CpG showed a TGI of 81.9% (P<0.001). Interestingly, the 26nt-CpG group exhibited faster weight gain, indicating better safety. In the mouse B16 melanoma model, LNP-delivered 26nt-CpG achieved a TGI rate of 75.2% (P<0.001), slightly higher than 30nt-CpG (71.0%, P<0.001), and also resulted in faster weight gain.

Conclusion: In conclusion, the use of LNP-delivered Toll-like receptor agonists demonstrated excellent tumor therapeutic effects and good safety in mouse models. Activation of Toll-like receptors effectively stimulated immune cells such as CD8+ T cells and NK cells, enhancing their ability to kill tumor cells. Direct injection of LNPs into the tumor improved this effect while reducing systemic side effects. This drug holds promise as a broad-spectrum anti-tumor treatment and may have potential for expanding the indications of existing tumor immunotherapies.

Citation Format: Jun Bai, Fei Su, Chen Li, Yujing Wu, Jing Li, Xiaobin Zhao, Yongsheng Yang, Robert J Lee. Lipid nanoparticle-delivered toll-like receptor agonists are highly effective in cancer immunotherapy [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr B049.
Abstract PR03: Hypoxia promotes inflammatory fibroblast formation in pancreatic cancer
Authors: Mello Ashley M. , Ngodup Tenzin , Lee Yusoo , Donahue Katelyn L. , Li Jinju , Rao Arvind , Carpenter Eileen S. , Crawford Howard C. , Magliano Marina Pasca Di , Lee Kyoung Eun
Published: December 2023

Pancreatic ductal adenocarcinoma (PDAC) is a deadly malignancy characterized by poor response to all existing therapies. Although immunotherapy has shown great promise against multiple deadly cancers, it has been largely ineffective in PDAC. This lack of response is in part attributed to its extensive, fibroinflammatory stroma and hypoxic microenvironment. Cancer-associated fibroblasts (CAFs) are a predominant and heterogeneous stromal cell type in PDAC, and single-cell transcriptomics of human and mouse PDAC has recently revealed a distinct CAF subpopulation, inflammatory CAFs (iCAFs). These inflammatory fibroblasts produce high levels of cytokines and chemokines in PDAC and have the potential to contribute to its immunosuppressive microenvironment and tumorigenesis. By injecting a hypoxia probe into PDAC mouse models, we recently found that iCAFs predominantly reside in hypoxic tumor regions. We also observed that the hypoxia-related gene signature is positively enriched in iCAFs in human PDAC samples. Importantly, by exposing three-dimensional (3D) co-cultures of pancreatic cancer cells and fibroblasts to either hypoxia or normoxia, we showed that hypoxia induces IL1α from cancer cells and that IL1α is required for hypoxia-mediated iCAF formation. Our data implicate hypoxia as a critical regulator of tumor stroma. Efforts are ongoing to understand the role of hypoxia in the crosstalk between cancer cells, fibroblasts, and immune cells in driving an immunosuppressive tumor microenvironment.

Citation Format: Ashley M. Mello, Tenzin Ngodup, Yusoo Lee, Katelyn L. Donahue, Jinju Li, Arvind Rao, Eileen S. Carpenter, Howard C. Crawford, Marina Pasca Di Magliano, Kyoung Eun Lee. Hypoxia promotes inflammatory fibroblast formation in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr PR03.
Abstract PR05: Dgat1 camouflages sexual dimorphism in anti-tumor CD8+ T cell immunity
Authors: Madi Alaa , Cui Guoliang
Published: December 2023

The tumor microenvironment (TME) is enriched in various lipid species that are detrimental to anti-tumor T cell function. However, intratumoral CD8+ T cells were recently shown to require free fatty acids, which are also abundant in the TME, for survival and function. It is not clear whether these free fatty acids regulate transcription or serve as an energy source either directly or through a triacylglyerol (TAG) intermediate step (as in memory CD8+ T cells). Here, we generated mice with T cell-specific deletion of Dgat1, the enzyme that catalyzes the last and rate limiting step in TAG synthesis. We found that the function of Dgat1 in melanoma-infiltrating CD8+ T cells is sexually dimorphic. In female mice, loss of Dgat1 results in much smaller tumors whereas in male mice Dgat1 deficiency limits anti-tumor T cell function. Mechanistically, Dgat1 is required for the survival of stem-cell like Tim3-Slamf6+CD8+ T cells in male mice, but actively decreases their abundance in female mice in a CD8+ T cell-intrinsic manner. Using a number of mouse models we reveal that androgen receptor signaling induces the accumulation of lipid peroxides and the activation of the unfolded protein response in Dgat1-deficient CD8+ T cells. Our study provides insights into a previously unexplored metabolic adaptation of tumor-infiltrating T cells to sex hormones.

Citation Format: Alaa Madi, Guoliang Cui. Dgat1 camouflages sexual dimorphism in anti-tumor CD8+ T cell immunity [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr PR05.
Supplementary Table S1 from CAR T-cell Design-dependent Remodeling of the Brain Tumor Immune Microenvironment Modulates Tumor-associated Macrophages and Anti-glioma Activity
Authors: Haydar Dalia , Ibañez-Vega Jorge , Crawford Jeremy Chase , Chou Ching-Heng , Guy Clifford S. , Meehl Michaela , Yi Zhongzhen , Perry Scott , Laxton Jonathan , Cunningham Trevor , Langfitt Deanna , Vogel Peter , DeRenzo Christopher , Gottschalk Stephen , Roussel Martine F. , Thomas Paul G. , Krenciute Giedre
Published: December 2023

Supplementary Table S1
Supplementary Figure 14 from CAR T-cell Design-dependent Remodeling of the Brain Tumor Immune Microenvironment Modulates Tumor-associated Macrophages and Anti-glioma Activity
Authors: Haydar Dalia , Ibañez-Vega Jorge , Crawford Jeremy Chase , Chou Ching-Heng , Guy Clifford S. , Meehl Michaela , Yi Zhongzhen , Perry Scott , Laxton Jonathan , Cunningham Trevor , Langfitt Deanna , Vogel Peter , DeRenzo Christopher , Gottschalk Stephen , Roussel Martine F. , Thomas Paul G. , Krenciute Giedre
Published: December 2023

Supplementary Figure S14 shows macrophage populations post CAR T-cell treatment in GL261 glioma bearing mice.
Avaliação diária de atitudes e profissionalismo na Graduação Médica: apresentação de um instrumento
Authors: Garcia Cristiane Pereira Pedro , Duarte Márcia Simei Zanovello , Duarte Sinésio Grace , Brunherotti Marisa Afonso de Andrade
Published: December 2023

Objetiva-se apresentar um instrumento de avaliação diária de atitudes e profissionalismo para estudantes de Medicina em atividades teórico-práticas. A elaboração do instrumento foi baseada nos manuais do programa de integração do aluno com a comunidade, projeto pedagógico do curso e nas Diretrizes Curriculares Nacionais da Graduaçãoem Medicina, realizada por docentes. Os docentes foram consultados em reuniões com duração de 50 minutos, com periodicidade semanal, entre agosto e novembro de 2016. Ao final do processo foi consolidada uma versão do instrumento com cinco itens e seis descritores para discriminação de situações de aprendizagem que permitem a avaliação da competência de um nível mais simples até o mais complexo. Com a utilização do instrumento elaborado não se deixa de avaliar pontos considerados importantes para a formação médica em atividades teórico-práticas.
A minha candidatura
Authors: Barreto Lima
Published: December 2023

Texto por Lima Barreto.
Não silenciou sobre o seu tempo
Authors: Lins Osman
Published: December 2023

Texto por Osman Lins.
Como introduzir com ordem, num espaço limitado, tudo o que pretendemos?: a importância do espaço na ficção osmaniana
Authors: Hazin Elizabeth
Published: December 2023

Partindo não apenas da observação do que Osman Lins deixou registrado em cartas, diários e em inúmeras entrevistas, mas também da certeza de que esse autor se dedicava com afinco à leitura de textos de teoria literária, o presente artigo pretende evidenciar sua preocupação com a construção de sua obra, enfatizando sobretudo as reflexões por ele imprimidas em Avalovara, a respeito da categoria “espaço literário”.
Apresentação
Authors: Nitrini Sandra , Hossne Andrea Saad
Published: December 2023

Apresentação por Andrea Saad Hossne e Sandra Nitrin.
Metodologias Ativas de Ensino-aprendizagem e Educação Especial: Relato de uma Experiência em Fisiologia da Audição no Ensino Superior
Authors: Oliveira Patricia de Sousa , Pereira-Costa Cyrce Leonor , Vigário Ana Flávia , Moreira Karla Graziella , Luiz-Ferreira Anderson
Published: December 2023

Nos últimos anos, temos observado um avanço nas políticas de acesso e permanência de jovens e adultos com necessidades educacionais especiais no Ensino Superior. A fim de atender a demanda destes alunos, professores têm buscado novas metodologias e materiais que possam facilitar os processos de ensino-aprendizagem. Neste contexto, o uso das metodologias ativas, ao colocar o estudante como centro das ações educativas e o conhecimento como resultado de um processo construído colaborativamente, tem se mostrado uma importante estratégia para a Educação Especial. A criação de ambientes e recursos, que possam ser explorados pelo estudante, possibilita interação, problematização e ressignificação do conhecimento. Diante disto, este trabalho teve como objetivo relatar o desenvolvimento de um recurso tátil e seu uso como metodologia ativa de ensino-aprendizagem de fisiologia da audição. Embora, movimentos mais dialógicos e interativos de aprendizagem possam ser desenvolvidos nas aulas, o ensino de fisiologia tradicionalmente se baseia no uso exclusivo de recursos audiovisuais, no qual alunos com deficiência visual não são assistidos adequadamente. Tal recurso foi confeccionado em biscuit, tendo suas principais estruturas envolvidas com a transdução auditiva contempladas. A utilização de materiais concretos adaptados permitiu aos estudantes explorar os conteúdos abordados, de forma a representar mentalmente o que foi tateado. Além disso, o modelo didático permitiu a interação entre os estudantes, a simulação do funcionamento do processo auditivo em condições normais e a problematização de contextos fisiológicos patológicos do sistema, possibilitando a participação ativa dos estudantes na construção do conhecimento.
Iron salt supplementation during gestation and gestational diabetes mellitus
Authors: Miranda Vanessa Iribarrem Avena , Pizzol Tatiane da Silva Dal , Silveira Marysabel Pinto Telis , Mengue Sotero Serrate , Silveira Mariângela Freitas da , Lutz Bárbara Heather , Bertoldi Andréa Dâmaso
Published: December 2023

OBJETIVE: To evaluate the association between the use of iron salts during the first two trimesters of gestation in non-anemic women and the development of gestational diabetes mellitus. METHODS: The study used maternal data from the 2015 Pelotas Birth Cohort. All non-anemic women at the 24th week of gestation (n = 2,463) were eligible for this study. Gestational diabetes mellitus was self-reported by women. Crude and adjusted logistic regression were performed considering level of significance = 0.05. RESULTS: Among the women studied, 69.7% were exposed to prophylactic iron supplementation in the first two trimesters of gestation. The prevalence of gestational diabetes mellitus among those exposed was 8.7% (95%CI: 7.4–10.1) and 9.3% (95%CI: 7.4–11.6) among those who were not exposed. Iron supplementation was not associated with increased risk of gestational diabetes mellitus in crude (OR = 0.9; 95%CI: 0,7–1,3) and adjusted analysis (OR = 1.1; 95%CI :0,8–1,6). CONCLUSIONS: The results suggest that routine iron use in non-anemic pregnant women does not increase the risk of developing gestational diabetes. This evidence supports the existing national and international guidelines, in which prophylactic iron supplementation is recommended for all pregnant women as soon as they initiate antenatal care in order to prevent iron deficiency anemia.
Soufisme et Chérifisme. La généalogie discutée d’une famille vénérée : les chorfa Baqqālī
Authors: Boudchar Mohamed Reda
Published: December 2023

The return of the chorfa, especially the Idrīsids, on the social and political scene in Morocco from the 14th century onwards, and the advantages and privileges they acquired thanks to their symbolic capital, led to a vast polemic on the authenticity of their prophetic descent, at a period that saw the emergence of many individuals and families claiming to be genealogically linked to the Prophet. This is why the Baqqālī family, of Idrīsid origin, suffered the rejection of their sharifian nasab, especially after having left the rural world for Tétouan, a city where other sharifian families had settled. The controversy grew between the intellectual elite and the people in charge of verification (ahl al-tawṯīq), and its echo touched historiography, since this sharifian family also represented Sufi sanctity, combining spiritual influence with elements of domination and social privilege.
Abū l-Hudà al-Ṣayyādī (1850-1909), the niqābat al-ašrāf, and Sufism between Sunnah and Šīʿah in Late Ottoman Iraq (Šarīfism in Modernity I)
Authors: Jordan David
Published: December 2023

During the late nineteenth and early twentieth century, the Ottoman state’s efforts to centralize the Syndicate of Prophetic Descendants (niqābat al-ašrāf) was paralleled by the massive growth of Prophetic descendants (sādah and ašrāf) and the mass conversion to the Šīʿah among the tribes in the Iraqi provinces. This contribution aims to shed new light on the role of Abū l-Hudà al-Ṣayyādī (1266/1850-1327/1909), the imperial naqīb al-ašrāf and paramount šayḫ and architect of the Sunnī Rifāʿiyyah Sufi order, in this context. Analyzing a selection of Ṣayyādī’s publications on genealogies, Sufi doctrine, and poem collections, the article focuses on his contribution to the emergence of a special subgroup of Prophetic descendants in the Arab tribal landscape, namely al-sādah al-rifāʿiyyah, and on the significance of his genealogical constructions for his understanding of Sunnī Islam as distinct from Šīʿism.
Quête des ansāb et quête des ašrāf dans le monde arabe (XVe-XXe siècle)
Authors: Mayeur-Jaouen Catherine
Published: December 2023

One hundred years after Lévi-Provençal’s Les Historiens des Chorfa and almost twenty-five years after Oriente Moderno’s founding issue on sādāt and ašrāf, a new special issue of Oriente Moderno is devoted to the “quest for ansāb”, along with the quest for ašrāf. Our project focuses mainly on Morocco, and on modern and contemporary sources. Firstly, we will take stock of twentieth- and twenty-first-century historiography, with works devoted to Arab genealogy (Zoltàn Szombathy) and “sayyido-sharifology” (Kazuo Morimoto), and to the intersection of the two (Sara Bowen Savant and Helena de Felipe). Modern and contemporary history is still relatively neglected in a field of research favored by medievalists and ethnologists. Our project aims to reflect on the links between genealogical science, the writing of history and the question of ašrāf, at a time of religious politics. imperial politics and the construction of modern states. Focusing on Morocco, this issue adopts a comparative spirit with three studies on Iraq, genealogical literature in Ottoman Egypt and contemporary Mauritania.
The Role of Gassendi’s Dutch Correspondents in his Astronomical Work
Authors: Pantin Isabelle
Published: December 2023

Gassendi was much engaged in observational astronomy and in the effort to circulate astronomical knowledge. He corresponded with astronomers living in several European countries. The exchanges he had with Dutch correspondents, notably Martin Hortensius and Govaart Wendelen, were among the most fruitful. These exchanges, which followed the rules of the Republic of Letters, circulated information in support of long-term projects, such as the revision of geographical coordinates and cosmic dimensions, at the moment when the use of the telescope in combination with traditional measuring instruments established a higher standard of accuracy.
An Analysis of the Effects of Clouds in High-Resolution Forecasting of Surface Shortwave Radiation in South Africa
Authors: Mendes Joana , Zwane Nosipho , Mabasa Brighton , Tazvinga Henerica , Walter Karen , Morcrette Cyril J. , Botai Joel
Published: December 2023

We assess site-specific surface shortwave radiation forecasts from two high-resolution configurations of the South African Weather Service numerical weather prediction model, at 4 and 1.5 km. The models exhibit good skill overall in forecasting surface shortwave radiation, with zero median error for all radiation components. This information is relevant to support a growing renewable energy sector in South Africa, particularly for photovoltaics. Further model performance analysis has shown an imbalance between cloud and solar radiation forecasting errors. In addition, cloud overprediction does not necessarily equate to underestimating solar radiation. Overcast cloud regimes are predicted too often with an associated positive mean radiation bias, whereas the relative abundance of partly cloudy regimes is underpredicted by the models with mixed radiation biases. Challenges highlighted by the misrepresentation of partly cloudy regimes in solar radiation error attribution may be used to inform improvements to the numerical core, namely, the cloud and radiation schemes.

Significance Statement

This paper provides the first comprehensive assessment of high-resolution site-specific NWP forecasts of surface shortwave radiation in South Africa, exploring clouds as the main drivers of prediction biases. Error attribution analyses of this kind are close to none for this part of the world. Our study contributes to understanding how cloud and radiation schemes perform over South Africa, representing a step forward in the state of the art. In addition to the scientific interest, the capabilities developed through this work may benefit the second largest economy of the continent. In a country where energy security is of critical relevance, the availability of useful and usable weather information is paramount to support its industry and socioeconomic growth.
Some Reflections on the 50th Anniversary of Hospice Care in America
Authors: Corr Charles A.
Published: December 2023

This article celebrates the 50th anniversary of hospice care in America. It also offers a brief recounting of one person’s reflections on some aspects of the history, achievements, and current status of American hospice care.
Global circulation of low-end expertise: Knowledge, hierarchy, and labor migration in a Burmese oilfield
Authors: Ren Chao
Published: December 2023

This article examines the phenomenon of the “global circulation of low-end expertise” through an exploration of the social dynamics surrounding American oil drillers who migrated from the Pennsylvania oil region to British colonial Burma during the early 1900s to the mid-1930s. These working-class drillers, with practical knowledge in oil drilling acquired through familial and community networks, played a crucial role in operating mechanized oil wells and providing geological expertise in colonial Burma. Positioned between labor-intensive agricultural economies in colonial Asia and the higher echelons of British colonial society, these drillers occupied an intermediate social location. Despite their indispensable expertise, they were marginalized due to their lower social standing, leading to their expertise being disregarded by their superiors and forgotten over time. By understanding the complexities of the “global circulation of low-end expertise,” this study sheds light on the social construction and erasure of the expertise held by these working-class drillers, revealing overlooked aspects of global histories of science and labor and highlighting the need to reassess dominant historical narratives on knowledge-labor.