Endogenous Response Research Articles

A core scientific problem in the treatment of central nervous system diseases: newborn neurons.

It has long been asserted that failure to recover from central nervous system diseases is due to the system's intricate structure and the regenerative incapacity of adult neurons. Yet over recent decades, numerous studies have established that endogenous neurogenesis occurs in the adult central nervous system, including humans'. This has challenged the long-held scientific consensus that the number of adult neurons remains constant, and that new central nervous system neurons cannot be created or renewed. Herein, we present a comprehensive overview of the alterations and regulatory mechanisms of endogenous neurogenesis following central nervous system injury, and describe novel treatment strategies that target endogenous neurogenesis and newborn neurons in the treatment of central nervous system injury. Central nervous system injury frequently results in alterations of endogenous neurogenesis, encompassing the activation, proliferation, ectopic migration, differentiation, and functional integration of endogenous neural stem cells. Because of the unfavorable local microenvironment, most activated neural stem cells differentiate into glial cells rather than neurons. Consequently, the injury-induced endogenous neurogenesis response is inadequate for repairing impaired neural function. Scientists have attempted to enhance endogenous neurogenesis using various strategies, including using neurotrophic factors, bioactive materials, and cell reprogramming techniques. Used alone or in combination, these therapeutic strategies can promote targeted migration of neural stem cells to an injured area, ensure their survival and differentiation into mature functional neurons, and facilitate their integration into the neural circuit. Thus can integration replenish lost neurons after central nervous system injury, by improving the local microenvironment. By regulating each phase of endogenous neurogenesis, endogenous neural stem cells can be harnessed to promote effective regeneration of newborn neurons. This offers a novel approach for treating central nervous system injury.

Max Share Identification of Multiple Shocks: An Application to Uncertainty and Financial Conditions

We generalize the Max Share approach to allow for simultaneous identification of a multiplicity of shocks in a Structural Vector Autoregression. Our machinery therefore overcomes the well-known drawbacks that individually identified shocks (i) tend to be correlated to each other or (ii) can be separated under orthogonalizations with weak economic ground. We show that identification corresponds to solving a nontrivial optimization problem. We provide conditions for non-emptiness of solutions and point-identification, and Bayesian algorithms for estimation and inference. We use the approach to study the effects of uncertainty and financial shocks, allowing for the possibility that the former responds contemporaneously to other shocks, distinguishing macroeconomic from financial uncertainty and credit supply shocks. Using U.S. data we find that financial uncertainty mimics a demand shock, while the interpretation of macro uncertainty is more mixed. Furthermore, variation in uncertainty partially represents the endogenous response of uncertainty to other shocks.

Abstract 4066: Generation of tumor targeted self-assembling split IL-12 subunits for the treatment of cancer

Abstract A major milestone in immuno-oncology was the development of immune checkpoint inhibitors, resulting in significant benefits for numerous patients. However, many patients are not responsive, become refractory, or cannot tolerate the treatment due to toxicity or side effects, demonstrating the need for additional immunotherapies. Cytokines are a class of promising immunomodulatory proteins being explored as therapeutics, but their success has been limited due to their rapid clearance or pleiotropic properties. Classically known as ‘signal 3’ of an endogenous immune response, IL-12p70 is a heterodimeric cytokine comprising p35 and p40 subunits. When administered exogenously, IL-12 is a potent stimulator of the immune system and can have profound anti-tumor activity; however, the clinical use of IL-12 has been limited due to poor systemic tolerability. Therefore, developing an immunotherapy that can be systemically administered and improve the therapeutic index could capitalize on the potential clinical benefit of IL-12. Mural Oncology has developed an innovative approach to mitigate the toxicity of IL-12 by splitting the heterodimer into inactive p35 and p40 monomers. The individual subunits are separately fused to two non-competitive antibody fragments targeting a highly expressed tumor-associated antigen. The goal of this approach is to conditionally activate IL-12 preferentially in the tumor microenvironment (TME). This is achieved by sequential administration of the targeted subunits, which will drive assembly and activity of the IL-12 heterodimer primarily at the tumor site, reducing systemic exposure and thereby potentially reducing associated toxicities. To engineer the split IL-12 subunits, a structure-based rational design approach was used to remove the natural intra-molecular disulfide bond and generate variants that maintained affinity between the p35 and p40 monomers. The engineered subunits were characterized using analytical methods and screened for the pair with the strongest affinity. Additionally, the candidate pair was assessed for receptor binding and functional activity on human T cells through phosphorylation of STAT4 and secretion of IFN-γ. Pharmacokinetics and pharmacodynamics were evaluated in mouse models with and without tumors demonstrating that dual targeting led to an increase in tumor retention, rapid clearance from circulation, and dose dependent release of IFN-γ. The results thus far demonstrate that the innovative approach of tumor targeted self-assembling split IL-12 has the potential to enhance the overall therapeutic potential of this cytokine. A development candidate will be identified and moved into clinical testing in cancer patients to enable us to fully understand if this design approach holds true in the clinical setting. Citation Format: Joshua Heiber, Pinar Gurel, Robert G. Newman, Kristiana Dreaden, Yanchun Zhao, Chunhua Wang, Su-Ping Pearson, Jean Chamoun. Generation of tumor targeted self-assembling split IL-12 subunits for the treatment of cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4066.

Abstract 7480: Human LINE-1 ORF1p retrotransposition independent function in pancreatic cancer

Abstract The aberrant expression of repeat elements is a common feature found in both mouse models and human pancreatic ductal adenocarcinoma (PDAC). The majority of these are non-coding RNAs that have shown the ability to stimulate a viral-like innate immune response. The long interspersed nuclear element 1 (LINE-1) retrotransposon is one of the few protein coding repeat elements in the human genome. LINE-1 has two open reading frames, ORF1 and ORF2. The former (ORF1p) encodes a ribonucleotide protein and the latter (ORF2p) a reverse transcriptase and endonuclease. Although nuclear ORF1p and ORF2p is required for retrotransposition, ORF1p is primarily localized in the cytoplasm, which suggests the possibility of ORF1p function independent of LINE-1 retrotransposition. Here, we show that ORF1p has a function to shield repeat RNAs from activating pathogen recognition receptor (PRR) mediated antiviral response in PDAC cells. LINE-1 ORF1p-targeting shRNA significantly inhibits patient-derived PDAC tumor growth both in vitro and in vivo. Total RNA sequencing (RNA-seq) reveals distinct transcriptional profiles with ORF1p deficiency. Gene set enrichment analysis (GSEA) identified high enrichment of innate immune response-related pathways including IL6-JAK-STAT3 signaling, inflammatory response, interferon response, and apoptosis in ORF1 knockdown PDAC cell lines. These immune related pathways are suppressed by deletion of RIG-I and MAVS, two components of viral RNA sensing. Mechanistically, we demonstrated that ORF1p is associated with repeat RNAs including LINE-1 RNA (L1HS) in processing bodies (PBs), which are cytoplasmic membraneless ribonucleoprotein (RNP) granules. In the PBs, ORF1p interacts with MOV10 RNA helicase, an essential enzyme for dsRNA unwinding. Using MOV10 truncated mutants, we found that the ORF1p-MOV10 interaction is important in attenuation of PRR sensing and apoptosis upon repeat RNA exposure. Finally, we evalulated the activation of the innate immune response in the tumor microenvironment by ORF1p depletion. These results suggest that ORF1p loss induces a desmoplastic reaction driven by Acta2+ myofibroblastic CAFs in response to inflammatory cytokines. Collectively, our results demonstrate a critical and previously uncharacterized role of ORF1p in evasion of endogenous repeat RNA-triggered antiviral immune response in PDAC cells with implications in the tumor microenvironment. Citation Format: Eunae You, Bidish Patel, Alexandra Rojas, Natalie Ho, Siyu Sun, Michael Raabe, Yuhui Song, Ildiko Phillips, Katherine Xu, Peter Richieri, Linda Nieman, Benjamin Greenbaum, David Ting. Human LINE-1 ORF1p retrotransposition independent function in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7480.

Abstract 4216: Indication agnostic evaluation of CAR T cell function using patient-derived immunocompetent tumoroid modelling platform

Abstract Introduction: Engineered T cells redirected towards tumor-specific cytotoxicity is a promising immunotherapeutic modality that has been well characterized for efficacy against B cell malignancies and evaluated in multiple tumor types. However, the development and optimization of CAR T cell therapies requires a rigorous preclinical model for the evaluation of CAR T cell function, tumor killing, cytokine production, and memory responses. Experimental Procedure: We are evaluating our model platform Custom Organoid Modelling Platform for Accurate and Speedy SolutionS (COMPASS 2) as a comprehensive platform for testing developmental and functional stages of CAR T cells including activation, memory, and exhaustion. We are performing qualitative and quantitative analyses of surface markers to recapitulate the complexity of human cancer. Additionally, since CAR T cells mediate crosstalk with other immune components in the TME, we also evaluate and monitor the presence of B cells, NK cells, macrophages, and endogenous T cells. Triple Negative Breast Cancer (TNBC); Immunophenotyping (IP); Immunofluorescence (IF); Fluorescence Activated Cell Sorting (FACS); T regulatory (Treg). Summary of Results: Our evaluation of COMPASS 2 for functional testing of CAR T cells demonstrates the feasibility of the platform in preclinical CAR T cell evaluation. We showed T cell infiltration and analyzed cytokine production that may alter tumor cells and TME. Crosstalk with other immune cells will be evaluated. We are currently evaluating the tumor volume changes using an EGFRvIII-targeted CART cell therapy on TNBC model systems, followed by apoptosis and EGFR analysis using FACS. Conclusion: Our COMPASS 2 model platform enables studies that require recapitulating hurdles to CAR therapy, including antigen escape, T cell dysfunction and trafficking; to reveal an essential role of the endogenous immune response mediating the antitumor effect following CAR therapy. Citation Format: Damieanus O. Ochola, Huckie Dell Mundo, Preethi Samuel, Natalia Korkorina, Bhuvanesh Dave. Indication agnostic evaluation of CAR T cell function using patient-derived immunocompetent tumoroid modelling platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4216.

Abstract 6238: Integrated multi-omic/modal profiling enables improved characterization of oncogenic processes, with impact to clinical actionability in the GenMineTOP tested population

Abstract Introduction: It is becoming increasingly apparent that tumor gene expression and spatial H&E profiling can be used to improve the accuracy of DNA-only tumor profiling for clinically relevant use-cases. Future development and deployment of integrative (multi-omic (DNA+RNA) and/or multi-modality (omics + imaging)) tests for treatment and clinical trial profiling hinges on the demonstration of clear robustness and performance gains of prognostic and predictive integrative predictors in a clinical setting. In this work we present several analyses quantifying the utility of integrative testing. We show that a multi-omic test combining tumor expression signatures with DNA profiles substantially improves tumor-of-origin prediction for cancers of unknown primary and also significantly improves MSI detection and resolution. We also show that a simple multi-modality model combining spatial features of tumor infiltrating lymphocytes of tumors with tumor expression is prognostic for MSI-positive patients. Methods: Multi-omicTumor-of-origin: 163 patient sample expression profiles were profiled using the GenMineTOP test. GenMineTOP is a tumor-normal, joint DNA+RNA capture PMDA-approved test in current clinical service in Japan. The samples were clustered using DNA-only, RNA-only, and joint DNA+RNA profiling signatures. MSI prediction: 940 samples from the TCGA COAD, UCEC, and READ cohorts with matched MMR IHC MSI-calling, DNA-NGS-based MSI calling, and RNA-seq data were utilized to train, test and validate an expression-based pan-cancer MSI caller. Additional orthogonal validation was carried out using GenMineTOP multi-omic profiles. Multi-modality Spatial context of TIL and tumor-expression: 158 MSI+ samples from the TCGA-UCEC cohort with matched fresh-frozen H&E 20X images and RNA-seq data were utilized to mine spatial features of tumor-infiltrating lymphocytes and derive gene expression signatures, respectively. A Cox-proportional hazard model for patient survival using gene expression and spatial TIL features was constructed while regressing out tumor stage, gender, age, and ethnicity. Results & Conclusions: Our multi-omic results quantify the benefit of combining DNA and RNA profiles from GenMineTOP to classify tumor-of-origin from unknown primaries reduces classification error by approximately 50%. Additionally, our expression-based MSI status prediction model not only significantly improves NGS DNA (MSI-sensor) based MSI prediction (log-ratio test, pval < 1e-7) but also strongly suggests that nonlinear differential expression drives the improvement in MSI prediction. Our multi-modality results demonstrate that mining the spatial context of endogenous tumor immune response has nontrivial prognostic utility (p < .05). Citation Format: Bojan Losic, Kenji Tatsuno, Vasanth R. Singan, Hiroki Ueda, Aya Shinozaki Ushiku, Vasanthan Jayakumar, Emi Hattori, Toshimitsu Ichijo, Takuma Sasa, Masakazu Akahori, Shinya Hayashi, Esther Hsiao, Stephanie Feupe, Dan Vo, Tiantian Geier, Shuichi Tsutsumi, Tetsuo Ushiku, Kousuke Watanabe, Katsutoshi Oda, Hidenori Kage, Joy Radecki-Crandall, Donavan T. Cheng, Hiroyuki Aburatani. Integrated multi-omic/modal profiling enables improved characterization of oncogenic processes, with impact to clinical actionability in the GenMineTOP tested population [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6238.

Targeting Epstein-Barr virus in multiple sclerosis: when and how?

Epidemiological evidence implicates Epstein-Barr virus (EBV) as the cause of multiple sclerosis (MS). However, its biological role in the pathogenesis of MS is uncertain. The article provides an overview of the role of EBV in the pathogenesis of MS and makes a case for targeting EBV as a treatment strategy for MS. EBV potentially triggers autoimmunity via molecular mimicry or immune dysregulation. Another hypothesis, supported by immunological and virological data, indicates that active EBV infection via latent-lytic infection cycling within the central nervous system or periphery drives MS disease activity. This supports testing small molecule anti-EBV agents targeting both latent and lytic infection, central nervous system-penetrant B-cell therapies and EBV-targeted immunotherapies in MS. Immunotherapies may include EBV-specific cytotoxic or chimeric antigen receptors T-cells, therapeutic EBV vaccines and immune reconstitution therapies to boost endogenous EBV-targeted cytotoxic T-cell responses. EBV is the probable cause of MS and is likely to be driving MS disease activity via latent-lytic infection cycling. There is evidence that all licensed MS disease-modifying therapies target EBV, and there is a compelling case for testing other anti-EBV strategies as potential treatments for MS.

Knockdown of orthotospovirus-derived silencing suppressor gene by plant-mediated RNAi approach induces viral resistance in tomato

RNA interference (RNAi) shows significant effectiveness in conferring resistance to viral infections in various plants. The current study was conducted to develop transgenic tomato (Solanum lycopersicum cv. Rio Grande) lines expressing a 320 bp conserved inverted region of the integral 35S promoter region. The molecular cloning was done for the construction of expression vectors in a hairpin structure by the Tomato spotted wilt orthotospovirus (TSWV) species Orthotospovirus tomatomaculae, silencing suppressor (NSs) gene. It contains both sense and antisense orientation in a binary vector pFGC5941. Short hairpin RNA (shRNA) structures were employed to reduce the possibility of inducing endogenous non-specific antiviral responses, which usually target longer double-stranded RNAs (dsRNAs). To confirm the transgenic plants, molecular analysis was performed using TSWV gene-specific primers (TSWV-NSs). Based on the EHA105 Agrobacterium strain and the pFGC5941 vector carrying NSs and nptII genes in the T-DNA region, the results validate the insertion of kanamycin resistance into the regenerated transgenic plants. The virus source harboring TSWV was used to inoculate putative transgenic plants for evaluating the confirmation of successful transformation strategy for inducing virus resistance. Quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed by using qRT-NSs primer pairs to determine the relative gene expression of all constructs (TSWV-NSs both sense and antisense orientation) after inoculations. In comparison to the control groups, which contained wild type control (non-transgenic tomato plant), empty pFGC5941 vector and the pFGC5941+sense construct, the expression of the NSs gene was noticeably reduced in the full clone (pFGC5941+sense + antisense) construct. For the validation of RNAi effectiveness, Nicotiana tabacum plants were also inoculated with viral constructs. Specifically, they displayed severe symptoms of TSWV in plants containing the pFGC5941+sense, pFGC5941 empty vector, and in wild-type tomato plants that are correlated with bioassay results. The lack of TSWV symptoms validated the hypothesis after careful observation of the expression of NSs genes in terms of pathogenicity. This research demonstrates the RNA interference effectiveness by targeting NSs gene as a reliable technique for achieving long lasting antiviral protection in tomatoes. The findings have practical relevance and the potential to be a tool for limiting tomato crop losses caused by TSWV worldwide.

Widespread S-persulfidation in activated macrophages as a protective mechanism against oxidative-inflammatory stress

Acute inflammatory responses often involve the production of reactive oxygen and nitrogen species by innate immune cells, particularly macrophages. How activated macrophages protect themselves in the face of oxidative-inflammatory stress remains a long-standing question. Recent evidence implicates reactive sulfur species (RSS) in inflammatory responses; however, how endogenous RSS affect macrophage function and response to oxidative and inflammatory insults remains poorly understood. In this study, we investigated the endogenous pathways of RSS biogenesis and clearance in macrophages, with a particular focus on exploring how hydrogen sulfide (H2S)-mediated S-persulfidation influences macrophage responses to oxidative-inflammatory stress. We show that classical activation of mouse or human macrophages using lipopolysaccharide and interferon-γ (LPS/IFN-γ) triggers substantial production of H2S/RSS, leading to widespread protein persulfidation. Biochemical and proteomic analyses revealed that this surge in cellular S-persulfidation engaged ∼2% of total thiols and modified over 800 functionally diverse proteins. S-persulfidation was found to be largely dependent on the cystine importer xCT and the H2S-generating enzyme cystathionine γ-lyase and was independent of changes in the global proteome. We further investigated the role of the sulfide-oxidizing enzyme sulfide quinone oxidoreductase (SQOR), and found that it acts as a negative regulator of S-persulfidation. Elevated S-persulfidation following LPS/IFN-γ stimulation or SQOR inhibition was associated with increased resistance to oxidative stress. Upregulation of persulfides also inhibited the activation of the macrophage NLRP3 inflammasome and provided protection against inflammatory cell death. Collectively, our findings shed light on the metabolism and effects of RSS in macrophages and highlight the crucial role of persulfides in enabling macrophages to withstand and alleviate oxidative-inflammatory stress.

High-sensitive detection of H2O2 in biological systems by persistent luminescent nanoprobes

Autofluorescence interference in the conventional fluorescence detection of hydrogen peroxide (H2O2) in biological systems represents a long-lasting problem in the field of biosensing. Here, we report a Cr3+, Y3+ co-doped ZnGaSnO4 (ZGSCY) nanoprobes which show long persistent luminescence (PL) that can be specifically enhanced by H2O2 without background fluorescence interference. The mesoporous organosilica are coated on the surface of ZGSCY to enhance the biocompatibility of the nanoprobes. Importantly, PL intensity of the nanoprobes can be further improved, because of the specific response of tetrasulfide bond in organosilica to H2O2. ZGSCY@MON show high-sensitive detection of H2O2 in living cells and human serum with the advantages of high specificity, low detection limit (91 nM), and wide linear range (0.5–60 μM, 60–400 μM). Furthermore, the extraordinary detection capability of H2O2 enables that the successful differentiate normal breast cells from breast cancer cells and the highly sensitive determination of serum glucose and cholesterol. We also clarified that the endogenous response originates from the UV-induced the interfacial electron transfer on ZGSCY regulated by H2O2 and exogenous response stems from the elimination of the adsorption of PL from the organosilica by H2O2. These findings open up a new effective way for autofluorescence-free detection of H2O2 in biological systems.

Abstract B084: Triple checkpoint blockade, but not anti-PD1 alone, enhances the efficacy of engineered adoptive T cell therapy in advanced ovarian cancer

Abstract Background: Over 20,000 women are diagnosed with ovarian cancer annually, and more than half will die within 5 years. This rate has changed little in the last 30 years, highlighting the need for therapy innovation. Although immunotherapy has revolutionized cancer treatment, efforts to harness endogenous patient immune responses have yielded limited therapeutic activity in ovarian cancer patients. T cells engineered to express a T cell receptor (TCR) targeting proteins uniquely overexpressed in tumors have the potential to control tumor growth without toxicity. Mesothelin (Msln) is over-expressed in ovarian cancer, contributes to the malignant and invasive phenotype, and has limited expression in healthy cells, making it a candidate immunotherapy target in these tumors. Methods: The ID8VEGF mouse cell line was used to evaluate if T cells engineered to express a mouse Msln-specific high-affinity T cell receptor (TCRMsln) can kill ovarian cancer. Tumor-bearing mice were treated with TCRMsln T cells plus anti-PD-1, anti-Tim-3 or anti-Lag-3 checkpoint-blocking antibodies alone or in combination, ultimately targeting up to three inhibitory receptors simultaneously. Single-cell RNA-sequencing (scRNAseq) was used to profile the impact of combination checkpoint blockade on engineered T cells and the tumor microenvironment (TME). Results: In a disseminated ID8 tumor model, adoptively transferred TCRMsln T cells preferentially accumulated in established tumors, delayed ovarian tumor growth, and prolonged mouse survival. However, elements in the TME limited engineered T cell persistence and cytolytic function. Triple checkpoint blockade, but not single- or double-agent treatment, dramatically increased antitumor function by intratumoral TCRMsln T cells. scRNAseq of tumor-infiltrating cells revealed distinct transcriptome changes in engineered and endogenous T cells and myeloid-derived cells. Engineered T cells, when combined with triple checkpoint blockade, increased expression of genes associated with effector and memory gene signatures, including proliferation and metabolic function, and reduced expression of genes associated with exhaustion. Moreover, combining adoptive immunotherapy with triple checkpoint blockade significantly prolonged survival in the cohort of treated tumor-bearing mice, relative to mice that received TCRMsln T cells alone or with anti-PD1 or double-agent treatments. Conclusions: Inhibitory receptor/ligand interactions within the TME can dramatically reduce T cell function, suggesting tumor cells may upregulate the ligands for PD-1, Tim-3 and Lag-3 for protection from tumor-infiltrating lymphocytes. In an advanced ovarian cancer model, triple checkpoint blockade significantly improved engineered T cell function and outcomes in mice in a setting where single checkpoint blockade had no significant activity. These results suggest that disrupting multiple inhibitory pathways simultaneously, which can be more safely pursed in a cell intrinsic form through genetic engineering, may be necessary for improved efficacy in patients. Citation Format: Kristin G. Anderson, Yapeng Su, Madison G. Burnett, Breanna M. Bates, Magdalia L. Rodgers Suarez, Susan L. Ruskin, Valentin Voillet, Raphael Gottardo, Philip D. Greenberg. Triple checkpoint blockade, but not anti-PD1 alone, enhances the efficacy of engineered adoptive T cell therapy in advanced ovarian cancer [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr B084.

DNAJB1-PRKACA fusion neoantigens elicit rare endogenous T cell responses that potentiate cell therapy for fibrolamellar carcinoma

Fibrolamellar carcinoma (FLC) is a liver tumor with a high mortality burden and few treatment options. A promising therapeutic vulnerability in FLC is its driver mutation, a conserved DNAJB1-PRKACA gene fusion that could be an ideal target neoantigen for immunotherapy. In this study, we aim to define endogenous CD8 Tcell responses to this fusion in FLC patients and evaluate fusion-specific Tcell receptors (TCRs) for use in cellular immunotherapies. We observe that fusion-specific CD8 Tcells are rare and that FLC patient TCR repertoires lack large clusters of related TCR sequences characteristic of potent antigen-specific responses, potentially explaining why endogenous immune responses are insufficient to clear FLC tumors. Nevertheless, we define two functional fusion-specific TCRs, one of which has strong anti-tumor activity invivo. Together, our results provide insights into the fragmented nature of neoantigen-specific repertoires in humans and indicate routes for clinical development of successful immunotherapies for FLC.

Targeting refractory/recurrent neuroblastoma and osteosarcoma with anti-CD3×anti-GD2 bispecific antibody armed T cells

BackgroundThe survival benefit observed in children with neuroblastoma (NB) and minimal residual disease who received treatment with anti-GD2 monoclonal antibodies prompted our investigation into the safety and potential clinical benefits...

Unveiling the dance of commodity prices and the global financial cycle

We examine the impact of commodity price changes on the business cycles and capital flows in emerging markets and developing economies (EMDEs), distinguishing between their role as a source of shock and as a channel of transmission of global shocks. Our findings reveal that surges in export prices, triggered by commodity price shocks, boost domestic GDP, an effect further amplified by the endogenous decline of country spreads. However, the effects on capital flows appear muted. Shifts in U.S. monetary policy and global risk appetite drive the global financial cycle in EMDEs. Eased global credit conditions, attributed to looser U.S. monetary policy or lower global risk appetite, lead to a rise in export prices, higher output, a decrease in government borrowing costs, and stimulate greater capital flows. The endogenous response of export prices amplifies the output effects of a more accommodative U.S. monetary policy while country spreads magnify the impact of shifts in global risk appetite.

Physico-Chemical Responses of Alstroemeria spp. cv. Rebecca to the presence of Salicylic Acid and Sucrose in vase solution during postharvest life.

The primary challenge in the cut flower industry, specifically in the postharvest phase, is the short vase life of flowers. This issue, along with early leaf yellowing and perianth abscission, significantly diminishes the economic value of flowers due to their accelerated senescence. To tackle this, we conducted a factorial experiment on Alstroemeria cv. Rebecca, utilizing a completely randomized design with three replications. In this experiment the effects of varying concentrations of Salicylic acid (SA) (0, 1.5, and 3mM) and sucrose (SU) (0% and 3%) were investigated on the postharvest quality of leaves and florets, with systematic evaluations every three days throughout their vase life. This experiment revealed that the specific treatment combination of 1.5mM SA + 3% SU (T5) markedly improved various parameters, such as vase life, total chlorophyll content, membrane stability index, relative fresh weight, and water uptake of cut flowers. In our analysis, we observed that this preservative solution not only extended the vase life and enhanced water uptake but also effectively preserved total chlorophyll, mitigated the loss of fresh weight, and reduced membrane deterioration in petals. Additionally, our results showed an increase in the activities of catalase (CAT) and peroxidase (POD) enzymes, as well as total protein content, alongside a decrease in malondialdehyde (MDA) and hydrogen peroxide (H2O2) levels. Moreover, this study noted a decrease in microbial populations in solutions containing different concentrations of salicylic acid. Our research demonstrated that alstroemeria flowers maintained in a solution with 1.5mM SA + 3% SU exhibited a significantly prolonged vase life of up to 21days, in contrast to the 15days observed in control flowers kept in water. These results are highly beneficial for manufacturers in the cut flower industry, as they provide a viable method to substantially extend the vase life of cut flowers. Such an enhancement in flower longevity can lead to increased market value and customer satisfaction. Furthermore, the reduction in flower senescence and decay rates can contribute to decreased waste and greater efficiency in cut flower distribution and sales, offering a substantial advantage to manufacturers in this competitive market. The extended vase life and reduced senescence observed in alstroemeria flowers treated with 1.5mM SA and 3% SU are attributed to SA's role in enhancing endogenous defense responses and sucrose's function as an energy source, collectively improving water uptake, and delaying the natural decay process.

Freezing stress response of wild and cultivated chickpeas.

Chickpea is an economically and nutritionally important grain legume globally, however, cold stress has adverse effects on its growth. In cold countries, like Canada where the growing season is short, having cold stress-tolerant varieties is crucial. Crop wild relatives of chickpea, especially Cicer reticulatum, can survive in suboptimal environments and are an important resource for crop improvement. In this study, we explored the performance of eleven C. reticulatum wild accessions and two chickpea cultivars, CDC Leader and CDC Consul, together with a cold sensitive check ILC533 under freezing stress. Freezing tolerance was scored based on a 1-9 scale. The wild relatives, particularly Kesen_075 and CudiA_152, had higher frost tolerance compared to the cultivars, which all died after frost treatment. We completed transcriptome analysis via mRNA sequencing to assess changes in gene expression in response to freezing stress and identified 6,184 differentially expressed genes (DEGs) in CDC Consul, and 7,842 DEGs in Kesen_075. GO (gene ontology) analysis of the DEGs revealed that those related to stress responses, endogenous and external stimuli responses, secondary metabolite processes, and photosynthesis were significantly over-represented in CDC Consul, while genes related to endogenous stimulus responses and photosynthesis were significantly over-represented in Kesen_075. These results are consistent with Kesen_075 being more tolerant to freezing stress than CDC Consul. Moreover, our data revealed that the expression of CBF pathway-related genes was impacted during freezing conditions in Kesen_075, and expression of these genes is believed to alleviate the damage caused by freezing stress. We identified genomic regions associated with tolerance to freezing stress in an F2 population derived from a cross between CDC Consul and Kesen_075 using QTL-seq analysis. Eight QTLs (P<0.05) on chromosomes Ca3, Ca4, Ca6, Ca7, Ca8, and two QTLs (P<0.01) on chromosomes Ca4 and Ca8, were associated with tolerance to freezing stress. Interestingly, 58 DEGs co-located within these QTLs. To our knowledge, this is the first study to explore the transcriptome and QTLs associated with freezing tolerance in wild relatives of chickpea under controlled conditions. Altogether, these findings provide comprehensive information that aids in understanding the molecular mechanism of chickpea adaptation to freezing stress and further provides functional candidate genes that can assist in breeding of freezing-stress tolerant varieties.

P17/C18-ceramide-mediated mitophagy is an endogenous neuroprotective response in preclinical and clinical brain injury.

Repeat concussions (or repetitive mild traumatic brain injury [rmTBI]) are complex pathological processes consisting of a primary insult and long-term secondary complications and are also a prerequisite for chronic traumatic encephalopathy (CTE). Recent evidence implies a significant role of autophagy-mediated dysfunctional mitochondrial clearance, mitophagy, in the cascade of secondary deleterious events resulting from TBI. C18-ceramide, a bioactive sphingolipid produced in response to cell stress and damage, and its synthesizing enzyme (CerS1) are precursors to selective stress-mediated mitophagy. A transporter, p17, mediates the trafficking of CerS1, induces C18-ceramide synthesis in the mitochondrial membrane, and acts as an elimination signal in cell survival. Whether p17-mediated mitophagy occurs in the brain and plays a causal role in mitochondrial quality control in secondary disease development after rmTBI are unknown. Using a novel repetitive less-than-mild TBI (rlmTBI) injury paradigm, ablation of mitochondrial p17/C18-ceramide trafficking in p17 knockout (KO) mice results in a loss of C18-ceramide-induced mitophagy, which contributes to susceptibility and recovery from long-term secondary complications associated with rlmTBI. Using a ceramide analog with lipid-selenium conjugate drug, LCL768 restored mitophagy and reduced long-term secondary complications, improving cognitive deficits in rlmTBI-induced p17KO mice. We obtained a significant reduction of p17 expression and a considerable decrease of CerS1 and C18-ceramide levels in cortical mitochondria of CTE human brains compared with age-matched control brains. These data demonstrated that p17/C18-ceramide trafficking is an endogenous neuroprotective mitochondrial stress response following rlmTBI, thus suggesting a novel prospective strategy to interrupt the CTE consequences of concussive TBI.

Recent Developments and Challenges in Projecting the Impact of Crop Productivity Growth on Biodiversity Considering Market-Mediated Effects.

The effect of an increase in crop productivity (output per unit of inputs) on biodiversity is hitherto poorly understood. This is because increased productivity of a crop in particular regions leads to increased profit that can encourage expansion of its cultivated area causing land use change and ultimately biodiversity loss, a phenomenon also known as "Jevons paradox" or the "rebound effect". Modeling such consequences in an interconnected and globalized world considering such rebound effects is challenging. Here, we discuss the use of computable general equilibrium (CGE) and other economic models in combination with ecological models to project consequences of crop productivity improvements for biodiversity globally. While these economic models have the advantage of taking into account market-mediated responses, resource constraints, endogenous price responses, and dynamic bilateral patterns of trade, there remain a number of important research and data gaps in these models which must be addressed to improve their performance in assessment of the link between local crop productivity changes and global biodiversity. To this end, we call for breaking the silos and building interdisciplinary networks across the globe to facilitate data sharing and knowledge exchange in order to improve global-to-local-to-global analysis of land, biodiversity, and ecosystem sustainability.

Lymph Node-Targeted Vaccine Boosting of TCR T-cell Therapy Enhances Antitumor Function and Eradicates Solid Tumors.

T-cell receptor (TCR)-modified T-cell therapies have shown promise against solid tumors, but overall therapeutic benefits have been modest due in part to suboptimal T-cell persistence and activation in vivo, alongside potential tumor antigen escape. In this study, we demonstrate an approach to enhance the in vivo persistence and function of TCR T cells through combination with Amphiphile (AMP) vaccination including cognate TCR T peptides. AMP modification improves lymph node targeting of conjugated tumor immunogens and adjuvants, thereby coordinating a robust T cell-activating endogenous immune response. AMP vaccine combination with TCR T-cell therapy led to complete eradication and durable responses against established murine solid tumors refractory to TCR T-cell monotherapy. Enhanced antitumor efficacy was correlated with simultaneous in vivo invigoration of adoptively transferred TCR T cells and in situ expansion of the endogenous antitumor T-cell repertoire. Long-term protection against tumor recurrence in AMP-vaccinated mice was associated with antigen spreading to additional tumor-associated antigens not targeted by vaccination. AMP vaccination further correlated with pro-inflammatory lymph node transcriptional reprogramming and increased antigen presenting-cell maturation, resulting in TCR T-cell expansion and functional enhancement in lymph nodes and solid tumor parenchyma without lymphodepletion. In vitro evaluation of AMP peptides with matched human TCR T cells targeting NY-ESO-1, mutant KRAS, and HPV16 E7 illustrated the clinical potential of AMP vaccination to enhance human TCR T-cell proliferation, activation, and antitumor activity. Taken together, these studies provide rationale and evidence to support clinical evaluation of combining AMP vaccination with TCR T-cell therapies to augment antitumor activity.

Lipoxins A4 and B4 inhibit glial cell activation via CXCR3 signaling in acute retinal neuroinflammation

Lipoxins are small lipids that are potent endogenous mediators of systemic inflammation resolution in a variety of diseases. We previously reported that Lipoxins A4 and B4 (LXA4 and LXB4) have protective activities against neurodegenerative injury. Yet, lipoxin activities and downstream signaling in neuroinflammatory processes are not well understood. Here, we utilized a model of posterior uveitis induced by lipopolysaccharide endotoxin (LPS), which results in rapid retinal neuroinflammation primarily characterized by activation of resident macroglia (astrocytes and Müller glia), and microglia. Using this model, we observed that each lipoxin reduces acute inner retinal inflammation by affecting endogenous glial responses in a cascading sequence beginning with astrocytes and then microglia, depending on the timing of exposure; prophylactic or therapeutic. Subsequent analyses of retinal cytokines and chemokines revealed inhibition of both CXCL9 (MIG) and CXCL10 (IP10) by each lipoxin, compared to controls, following LPS injection. CXCL9 and CXCL10 are common ligands for the CXCR3 chemokine receptor, which is prominently expressed in inner retinal astrocytes and ganglion cells. We found that CXCR3 inhibition reduces LPS-induced neuroinflammation, while CXCR3 agonism alone induces astrocyte reactivity. Together, these data uncover a novel lipoxin–CXCR3 pathway to promote distinct anti-inflammatory and proresolution cascades in endogenous retinal glia.