Cell Counterparts Research Articles

Optimal performance of silicon nanowire solar cells under low sunlight concentration and their integration as bottom cells in III–V multijunction systems

Nanostructured silicon solar cells are designed to minimize costs through reduced material usage while enhancing power conversion efficiency via superior light trapping and shorter charge separation distances compared to traditional planar cells. This study identifies the optimal conditions for nanoimprinted silicon nanowire (SiNW) solar cells to achieve maximum efficiency under low sunlight concentration and evaluates their performance as bottom cells in III–V multijunction solar cell systems. The findings indicate that the SiNW solar cell reaches its peak performance at a concentration factor of 7.5 suns and a temperature of 40°C or lower. Specifically, the absolute conversion efficiency under these conditions is 1.05% higher than that under unconcentrated light. Compared to a planar silicon solar cell under identical conditions, the SiNW solar cell exhibits a 3.75% increase in conversion efficiency. Additionally, the SiNW single-junction solar cell, when integrated in series with a commercial lattice-matched InGaP/GaAs dual-junction solar cell, was tested under unconcentrated sunlight, specifically at one-sun, global air mass 1.5 condition, to assess its viability in one-sun multi-junction solar cell applications. The results suggest that a III–V upper subcell with a smaller active area than that of the SiNW subcell is optimal for maximizing current production, which is favorable to the cost reduction of the device. This hybrid configuration is particularly advantageous for terrestrial applications, such as electric vehicles, which demand lightweight, high-performance multijunction solar cell devices. Although the weight reduction of the characterized SiNW solar cell with a full silicon substrate compared to its planar solar cell counterpart is 1.8%, recommendations to increase this reduction to as much as 64.5% are discussed to conclude this paper.

(Invited) Non-Fullerene Acceptor in Organic Solar Cells Toward Improving Performance as Indoor Light Energy Harvester

Research in organic solar cells aims to develop new materials that improve efficiency. The inception of emerging non-fullerene acceptors (NFAs) by replacing the fullerene counterpart in organic solar cells (OSCs) has pushed the bulk-heterojunction based photovoltaics to achieve efficiencies around 18% [1-6]. In order to produce efficient OSCs, it is required to have donor and acceptor materials with matching absorption bands in the Vis-NIR range, high charge-carrier mobility, and a small energy offset to minimize voltage losses. Mainly, the development of Y7 NFA molecule has improved power conversion efficiencies by pairing selected polymer donors, such as PM6 [7, 8]. Nowadays, OSCs have attracted great attention for being used as indoor light energy harvesters. The organic materials in the active layer can have tuned optical band gaps, providing an absorption spectrum that matches with emission spectra of several indoor lights, e.g., halogen lights, LED (light-emitting diodes), and FLs (fluorescent lamp), which have different emitting spectra [9, 10].The use of OSCs for indoor light harvesting applications is promising for integration into low-power indoor electronic devices from microwatts to milliwatts, such as data transfer systems, alarm systems and distributed controls. Moreover, the OSCs have the possibility to be used with power wireless sensor nodes connected to the Internet of Things. At present, LEDs are the most popular low-light level sources which can be found in offices and homes. The intensity of the light emitted by an artificial light source, per unit area of a surface is called illuminance and measured in units of lux (lx). A level 500 lx is typically recommended for office environments and 200 lx for living room environments.In this research, we have been aiming at the performance and stability over time of organic solar cells under LED illumination using fullerene and non-fullerene acceptors. We have chosen as the indoor illumination source an LED lamp with a color temperature of 3000 K and color rendering index (CRI) > 80 following the protocols ISOS [11]. The LED lamp output illuminances were varied in the range from 200 to 1500 lx. We have used PM6 as polymer donor material due to their absorption spectrum and energy band gaps well-matched to the incident LED spectrum. In order to research the improvement of the performance of OSCs under indoor illumination we have compared PC70BM fullerene acceptor with ITIC-M, Y6-O, and Y7 non-fullerene acceptors. As a result, under 1000 lx illumination we achieved PCEs from ~ 12% using Y6-O, ~ 15% using PC70BM, ~16% using Y7 up to more than 18% efficiency using ITIC-M. The study is performed including optical and electrical characterization with the purpose of understanding the behavior and the transport processes taking place in the device. In future work, we focus on the stability of the encapsulated OSCs under continuous LED illumination following the agreed stability testing protocols to study which active layer turns out to be more stable under these conditions. Acknowledgements: This work was supported by the Ministerio de Ciencia, Innovación y Universidades (MICINN/FEDER) PDI2021-128342OB-I00, by the Agency for Management of University and Research Grants (AGAUR) ref. 2021-SGR-00739, and by the Catalan Institution for Research and Advanced Studies (ICREA) under the ICREA Academia Award. M. Ramírez-Como acknowledge to CONAHCYT ref. BPPA-20220624083033039-2364083.

Deciphering The Transcriptional Activities of Genes Coding For Adipokines and Their Receptors in Porcine Ex Situ-Protected Mesenchymal Stem Cells Undergoing Adipogenic Differentiation

Abstract The aim of this study was to investigate the impact of the source of mesenchymal stem cells (MSCs) and their adipogenic derivatives on the relative abundances (RAs) noticed for mRNA transcripts of the selected adipokines (adiponectin and leptin) and their receptors. MSCs were isolated from bone marrow (BM) and subcutaneous adipose tissue (AT ) samples collected post mortem from a total of four gilts (each at the age of 6 months). The stemness of the MSCs was proven via recognizing their abilities to differentiate into adipocytes, osteoblasts, and chondrocytes. By using real-time PCR (RT -qPCR), the quantitative levels of transcriptional activity pinpointed for the adiponectin (ADIPOQ), adiponectin receptor 1 (ADIPOR1), adiponectin receptor 2 (ADIPOR2), leptin (LEP), and leptin receptor (LEPR) genes were comparatively analyzed between non-differentiated AT -MSCs and BM-MSCs and their cell counterparts undergoing differentiation into adipocytes. A significantly higher RA for ADIPOR1 transcripts was identified in the AT-MSCs compared to the BM-MSCs and their adipogenic derivatives (P≤0.05). Moreover, the quantitative levels of LEPR transcripts were shown to increase significantly among adipocytes originating from differentiated BM-MSCs compared to undifferentiated BM-MSCs, AT -MSCs, and AT -MSC-derived adipocytes (P≤0.05). Nonetheless, there was no significant inter-group variability in the RAs of the ADIPOQ, ADIPOR2, or LEP mRNA transcripts (P>0.05). Cumulatively, the in vitro models focused on the identification and detailed exploration of transcriptomic signatures of undifferentiated pig BM- and AT -MSCs, and the molecular mechanisms underlying the adipogenic differentiation pathways of the above-indicated two distinct sources of stem cells were developed and optimized in the current investigation for the very first time. These ex vivo porcine models might confirm the expedited functional mobilization of ex situ-protected MSCs and their enhanced capacity to be transcriptionally reprogrammed into adipocytes due to physiopathological alterations in the expression profiles of adipokines and their receptors, which are prompted and progressed in obese and superobese women patients at the peri-pubertal period of ontogenesis. Finally, the models providing comprehensive molecular insights into ex situ-protected porcine BM- and AT -derived MSCs and their differentiated derivatives can be largely suitable for the biotechnologically assisted rescue and restitution of endangered representatives of rare native breeds of domestic pig.

Pro- and antitumorigenic functions of γδ T cells.

γδ T cells are a subset of T cells that are characterized by the expression of a TCR-γδ instead of a TCR-αβ. Despite being outnumbered by their αβ T cell counterpart in many tissues, studies from the last 20 years underline their important and non-redundant roles in tumor and metastasis development. However, whether a γδ T cell exerts pro- or antitumorigenic effects seems to depend on a variety of factors, many of them still incompletely understood today. In this review, we summarize mechanisms by which γδ T cells exert these seemingly contradictory effector functions in mice and humans. Furthermore, we discuss the current view on inducing and inhibiting factors of γδ T cells during cancer development.

The role of CD20+ T cells: Insights in human peripheral blood.

CD20+ T cells constitute a small subset of T cells. These are found among CD4+, CD8+, CD4+CD8+, CD4-CD8- T, and TCRγδ+ T cells, and have been poorly characterized. The aim of this study was to characterize peripheral blood (PB) CD20+ T cells and compare them to their PB CD20- T cell counterparts. PB from 17 healthy individuals was collected. The distribution of CD20+ T cells among maturation-associated T cells compartments (naïve, central memory, transitional memory, effector memory, and effector T cells), their polarization, activation status, and expression of immune-regulatory proteins were evaluated by flow cytometry. Their function was also assessed, by measuring IFN-γ, TNF-α, and IL-17 production. Compared with CD20- T cells, CD20+ T cells represent a higher proportion of transitional memory cells. Furthermore, CD20+ T cells display a proinflammatory phenotype, characterized by the expansion of Th1, Th1/17, and Tc1 cell subsets , associated to a high expression of activation (CD25) and exhaustion (PD-1) markers. In addition, the simultaneous production of the proinflammatory cytokines IFN-γ, TNF-α, and IL-17 was also detected in CD4+CD20+ T cells. Our results show that CD20+ T cells are phenotypically and functionally different from CD20- T cells, suggesting that these cells are a distinct subset of T cells.

Endotoxin tolerance and trained immunity: breaking down immunological memory barriers.

For decades, innate immune cells were considered unsophisticated first responders, lacking the adaptive memory of their T and B cell counterparts. However, mounting evidence demonstrates the surprising complexity of innate immunity. Beyond quickly deploying specialized cells and initiating inflammation, two fascinating phenomena - endotoxin tolerance (ET) and trained immunity (TI) - have emerged. ET, characterized by reduced inflammatory response upon repeated exposure, protects against excessive inflammation. Conversely, TI leads to an enhanced response after initial priming, allowing the innate system to mount stronger defences against subsequent challenges. Although seemingly distinct, these phenomena may share underlying mechanisms and functional implications, blurring the lines between them. This review will delve into ET and TI, dissecting their similarities, differences, and the remaining questions that warrant further investigation.

Chimeric antigen receptor (CAR) modified T Cells in acute myeloid leukemia: limitations and expectations.

Acute myeloid leukemia (AML) is an aggressive hematologic malignancy with a poor prognosis despite the advent of novel therapies. Consequently, a major need exists for new therapeutic options, particularly for patients with relapsed/refractory (R/R) AML. In recent years, it has been possible to individualize the treatment of a subgroup of patients, particularly with the emergence of multiple targeted therapies. Nonetheless, a considerable number of patients remain without therapeutic options, and overall prognosis remains poor because of a high rate of disease relapse. In this sense, cellular therapies, especially chimeric antigen receptor (CAR)-T cell therapy, have dramatically shifted the therapeutic options for other hematologic malignancies, such as diffuse large B cell lymphoma and acute lymphoblastic leukemia. In contrast, effectively treating AML with CAR-based immunotherapy poses major biological and clinical challenges, most of them derived from the unmet need to identify target antigens with expression restricted to the AML blast without compromising the viability of the normal hematopoietic stem cell counterpart. Although those limitations have hampered CAR-T cell therapy translation to the clinic, there are several clinical trials where target antigens, such as CD123, CLL-1 or CD33 are being used to treat AML patients showing promising results. Moreover, there are continuing efforts to enhance the specificity and efficacy of CAR-T cell therapy in AML. These endeavors encompass the exploration of novel avenues, including the development of dual CAR-T cells and next-generation CAR-T cells, as well as the utilization of gene editing tools to mitigate off-tumor toxicities. In this review, we will summarize the ongoing clinical studies and the early clinical results reported with CAR-T cells in AML, as well as highlight CAR-T cell limitations and the most recent approaches to overcome these barriers. We will also discuss how and when CAR-T cells should be used in the context of AML.

Role of germinal center and CD39highCD73+ B cells in the age-related tonsillar involution

BackgroundThe tonsils operate as a protection ring of mucosa at the gates of the upper aero-digestive tract. They show similarities with lymph nodes and participate as inductive organs of systemic and mucosal immunity. Based on the reduction of their size since puberty, they are thought to experience involution in adulthood. In this context, we have used tonsillar mononuclear cells (TMC) isolated from patients at different stages of life, to study the effect of ageing and the concomitant persistent inflammation on these immune cells.ResultsWe found an age-dependent reduction in the proportion of germinal center B cell population (BGC) and its T cell counterpart (T follicular helper germinal center cells, TfhGC). Also, we demonstrated an increment in the percentage of local memory B cells and mantle zone T follicular helper cells (mTfh). Furthermore, younger tonsils rendered higher proportion of proliferative immune cells within the freshly isolated TMC fraction than those from older ones. We demonstrated the accumulation of a B cell subset (CD20+CD39highCD73+ cells) metabolically adapted to catabolize adenosine triphosphate (ATP) as patients get older. To finish, tonsillar B cells from patients at different ages did not show differences in their proliferative response to stimulation ex vivo, in bulk TMC cultures.ConclusionsThis paper sheds light on the changing aspects of the immune cellular landscape, over the course of time and constant exposure, at the entrance of the respiratory and digestive systems. Our findings support the notion that there is a re-modelling of the immune functionality of the excised tonsils over time. They are indicative of a transition from an effector type of immune response, typically oriented to reduce pathogen burden early in life, to the development of an immunosuppressive microenvironment at later stages, when tissue damage control gets critical provided the time passed under immune attack. Noteworthy, when isolated from such histologic microenvironment, older tonsillar B cells seem to level their proliferation capacity with the younger ones. Understanding these features will not only contribute to comprehend the differences in susceptibility to pathogens among children and adults but would also impact on vaccine developments intended to target these relevant mucosal sites.

Abstract SY45-02: Engineering strategies based on receptors derived from gdT cells

Abstract SY45-02: Engineering strategies based on receptors derived from gdT cells

Abstract LB062: ADT3_A potential first-in-class Vδ1-engager for in situ activation of Vδ1 γδ T cells and enhanced cytotoxicity towards solid tumors

Abstract Gamma delta (γδ) T cells are crucial in stress surveillance for infections and cancer. Studies have shown that tumour-infiltrating γδ T cells are a positive prognostic factor in cancer therapy. While γδ T cells share many characteristics with their αβ T cell counterpart, such as cytotoxic effector functions, they express a distinct TCR that is composed of a γ and a δ chain, belong to the HLA-class-I unrestricted immune cell subset with the ability to discriminate and specifically kill tumor cells via recognition of tumor stress ligands. Clinical trials using Vδ1 γδ T cell therapies have demonstrated promising results in Ph 1 trials, with early signs of favorable efficacy and safety profile.ADT3, is a novel bispecific engager that activates tissue-resident Vδ1+ T cells via (Epidermal Growth Factor Receptor) EGFR crosslinking on tumor cells. VD1 T cells isolated from skin biopsies, when activated, enhance the selective killing of EGFR positive tumor cells through innate mechanisms that recognize stress ligands that are otherwise absent on healthy cells. As a bispecific engager, ADT3 preserves high affinity binding to VD1 TCR through the Fab end of the molecule, and binding to two EGFR binding sites via the modified Fc domain. Crosslinking of VD1 and EGFR+ tumor cells with ADT3, mediates a series of events leading to proliferation of VD1 cells, upregulation of activation markers, CD107a degranulation and secretion of chemokines and cytokines (such as TNFa and IFNg) and tumor kill as determined by its potent cytotoxic activity in a T-cell dependent cellular cytotoxicity assay (TDCC). Apart from direct tumor kill, the secretion of such cytokines and chemokines via ADT3 activation are required for induction of immune orchestration of non-Vδ1 immune cells to enhance tumor kill. Preclinical data supports future clinical entry of ADT3, and IND enabling studies are in progress Citation Format: Joshua Freedman, Georgia Stevens, Julie Julie Hoong, Mihirban Tuna, Yaoyao Shi, Kathy J. Seidl, Katherine Galvin, Andrew Hutton, Antara Banerjee. ADT3_A potential first-in-class Vδ1-engager for in situ activation of Vδ1 γδ T cells and enhanced cytotoxicity towards solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB062.

The MDM2-p53 Axis Represents a Therapeutic Vulnerability Unique to Glioma Stem Cells.

The prevention of tumor recurrence by the successful targeting of glioma stem cells endowed with a tumor-initiating capacity is deemed the key to the long-term survival of glioblastoma patients. Glioma stem cells are characterized by their marked therapeutic resistance; however, recent evidence suggests that they have unique vulnerabilities that may be therapeutically targeted. We investigated MDM2 expression levels in glioma stem cells and their non-stem cell counterparts and the effects of the genetic and pharmacological inhibition of MDM2 on the viability of these cells as well as downstream molecular pathways. The results obtained showed that MDM2 expression was substantially higher in glioma stem cells than in their non-stem cell counterparts and also that the inhibition of MDM2, either genetically or pharmacologically, induced a more pronounced activation of the p53 pathway and apoptotic cell death in the former than in the latter. Specifically, the inhibition of MDM2 caused a p53-dependent increase in the expression of BAX and PUMA and a decrease in the expression of survivin, both of which significantly contributed to the apoptotic death of glioma stem cells. The present study identified the MDM2-p53 axis as a novel therapeutic vulnerability, or an Achilles' heel, which is unique to glioma stem cells. Our results, which suggest that non-stem, bulk tumor cells are less sensitive to MDM2 inhibitors, may help guide the selection of glioblastoma patients suitable for MDM2 inhibitor therapy.

Abstract 5233: Engineered NK-like NKG2C+ CD8 T cells mediate superior anti-tumor efficacy over conventional CD8 T cells and NK cells

Abstract We have recently characterized a unique CD8 T cell subset that expresses NKG2C, as well as multiple other natural killer cell markers. These NKG2C+ CD8 T cells demonstrate NK-like effector functions against various tumor targets, can be efficiently expanded and engineered using GMP-compatible techniques, and most importantly, display beneficial qualities unique to T and NK cells while minimizing the therapeutic deficits of each. NKG2C+ CD8 cells can be easily expanded using a genetically modified K562 feeder line and can be efficiently engineered via retroviral transduction. We genetically engineered NKG2C+ CD8 T cells to express an anti-CD19-28-z CAR construct to target B cell malignancies and compared effector functions of the CAR-engineered NKG2C+ CD8 T cells against CAR+NKG2C- (conventional) CD8 T cells and CAR+ NK cells in vitro and in vivo. We examined expansion, engineering efficiency, tumor control, and extent of cytokine release syndrome (CRS). We also engineered NKG2C+ CD8 T cells with a transgenic NY-ESO-1 TCR to target melanoma and compared the functional responses and efficacy of TCR-engineered NKG2C+ CD8 T cells against TCR-engineered conventional CD8 T cells. CAR+NKG2C+ CD8 T cells demonstrated superior in vitro effector function when compared to CAR+ conventional CD8 T cells and CAR+NK cells and did not upregulate characteristic T cell exhaustion markers, such as PD-1, TIM-3, and LAG-3. These CAR+NKG2C+ CD8 T cells demonstrated superior control of a CD19+ NALM6 tumor model in NRG mice, which was further enhanced with administration of IL-15. CAR+NKG2C+ CD8 T cells with IL-15 demonstrate dramatic and enhanced persistence, as well as superior cytotoxicity and activation in vivo when compared to CAR+ conventional CD8 T cells and CAR+ NK cells. Using a SCID-beige CRS mouse model, we observed reduced CRS in mice treated with CAR+ NKG2C+ CD8 T cells than in mice treated with CAR+ conventional CD8 T cells, with lower levels of proinflammatory cytokines and minimal expansion of murine myeloid subsets. Similarly, TCR-engineered NKG2C+ CD8 T cells demonstrated enhanced tumor control and in vivo persistence in an A375 melanoma-bearing NRG mouse model compared to TCR-engineered conventional CD8 T cells. We have successfully demonstrated that CAR- or TCR-engineered NKG2C+ CD8 T cells have superior anti-tumor efficacy, in vitro and in vivo, to conventional CD8 T cell and NK cell counterparts due to their innate tumor killing and greater in vivo persistence. These cells mediate less CRS and provide a safer alternative to traditional engineered T cell therapeutics. Ultimately, our NKG2C+ CD8 T cells represent a unique platform for engineered T cell therapies to target both solid tumors and hematologic malignancies, that is easily expanded ex vivo, does not require CD4 T cells for in vivo persistence, and is resistant to exhaustion. Citation Format: Kyle Lupo, M. Kazim Panjwani, Anthony Daniyan, Christopher Klebanoff, Katharine Hsu. Engineered NK-like NKG2C+ CD8 T cells mediate superior anti-tumor efficacy over conventional CD8 T cells and NK cells [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 5233.

IFNα induces CCR5 in CD4+ T cells of HIV patients causing pathogenic elevation

BackgroundAmong people living with HIV, elite controllers (ECs) maintain an undetectable viral load, even without receiving anti-HIV therapy. In non-EC patients, this therapy leads to marked improvement, including in immune parameters, but unlike ECs, non-EC patients still require ongoing treatment and experience co-morbidities. In-depth, comprehensive immune analyses comparing EC and treated non-EC patients may reveal subtle, consistent differences. This comparison could clarify whether elevated circulating interferon-alpha (IFNα) promotes widespread immune cell alterations and persists post-therapy, furthering understanding of why non-EC patients continue to need treatment.MethodsLevels of IFNα in HIV-infected EC and treated non-EC patients were compared, along with blood immune cell subset distribution and phenotype, and functional capacities in some cases. In addition, we assessed mechanisms potentially associated with IFNα overload.ResultsTreatment of non-EC patients results in restoration of IFNα control, followed by marked improvement in distribution numbers, phenotypic profiles of blood immune cells, and functional capacity. These changes still do not lead to EC status, however, and IFNα can induce these changes in normal immune cell counterparts in vitro. Hypothesizing that persistent alterations could arise from inalterable effects of IFNα at infection onset, we verified an IFNα-related mechanism. The protein induces the HIV coreceptor CCR5, boosting HIV infection and reducing the effects of anti-HIV therapies. EC patients may avoid elevated IFNα following on infection with a lower inoculum of HIV or because of some unidentified genetic factor.ConclusionsEarly control of IFNα is essential for better prognosis of HIV-infected patients.

Unlocking the power of sugar: carbohydrate ligands as key players in nanotherapeutic-assisted targeted cancer therapy.

Cancer cells need as much as 40-times more sugar than their normal cell counterparts. This sugar demand is attained by the excessive expression of inimitable transporters on the surface of cancer cells, driven by their voracious appetite for carbohydrates. Nanotechnological advances drive research utilizing ligand-directed therapeutics and diverse carbohydrate analogs. The precise delivery of these therapeutic cargos not only mitigates toxicity associated with chemotherapy but also reduces the grim toll of mortality and morbidity among patients. This in-depth review explores the potential of these ligands in advanced cancer treatment using nanoparticles. It offers a broader perspective beyond the usual ways we deliver drugs, potentially changing the way we fight cancer.

High and selective cytotoxicity of ex vivo expanded allogeneic human natural killer cells from peripheral blood against bladder cancer: implications for natural killer cell instillation after transurethral resection of bladder tumor

BackgroundNon-muscle-invasive bladder cancer (NMIBC) is treated with transurethral resection of bladder tumor (TURBT) followed by intravesical instillation of chemotherapy or Bacillus Calmette–Guérin therapy. However, these treatments have a high recurrence rate and side effects, emphasizing the need for alternative instillations. Previously, we revealed that expanded allogeneic human natural killer (NK) cells from peripheral blood are a promising cellular therapy for prostate cancer. However, whether NK cells exhibit a similar killing effect in bladder cancer (BCa) remains unknown.MethodsExpansion, activation, and cryopreservation of allogeneic human NK cells obtained from peripheral blood were performed as we previously described. In vitro cytotoxicity was evaluated using the cell counting kit-8. The levels of perforin, granzyme B, interferon-γ, tumor necrosis factor-α, and chemokines (C-C-motif ligand [CCL]1, CCL2, CCL20, CCL3L1, and CCL4; C-X-C-motif ligand [CXCL]1, CXCL16, CXCL2, CXCL3, and CXCL8; and X-motif ligand 1 and 2) were determined using enzyme-linked immunosorbent assay. The expression of CD107a, major histocompatibility complex class I (MHC-I), MHC-I polypeptide-related sequences A and B (MICA/B), cytomegalovirus UL16-binding protein-2/5/6 (ULBP-2/5/6), B7-H6, CD56, CD69, CD25, killer cell Ig-like receptors (KIR)2DL1, KIRD3DL1, NKG2D, NKp30, NKp46, and CD16 of NK cells or BCa and normal urothelial cells were detected using flow cytometry. Cytotoxicity was evaluated using lactate dehydrogenase assay in patient-derived organoid models. BCa growth was monitored in vivo using calipers in male NOD-scid IL2rg−/− mice subcutaneously injected with 5637 and NK cells. Differential gene expressions were investigated using RNA sequence analysis. The chemotaxis of T cells was evaluated using transwell migration assays.ResultsWe revealed that the NK cells possess higher cytotoxicity against BCa lines with more production of cytokines than normal urothelial cells counterparts in vitro, demonstrated by upregulation of degranulation marker CD107a and increased interferon-γ secretion, by MICA/B/NKG2D and B7H6/NKp30-mediated activation. Furthermore, NK cells demonstrated antitumor effects against BCa in patient-derived organoids and BCa xenograft mouse models. NK cells secreted chemokines, including CCL1/2/20, to induce T-cell chemotaxis when encountering BCa cells.ConclusionsThe expanded NK cells exhibit potent cytotoxicity against BCa cells, with few toxic side effects on normal urothelial cells. In addition, NK cells recruit T cells by secreting a panel of chemokines, which supports the translational application of NK cell intravesical instillation after TURBT from bench to bedside for NMIBC treatment.

The Extended Cleavage Specificity of Channel Catfish Granzyme-like II, A Highly Specific Elastase, Expressed by Natural Killer-like Cells.

The extended cleavage specificity of catfish granzyme-like II has been characterized using substrate phage display. The preference for particular amino acids at and surrounding the cleavage site was further validated by using a panel of recombinant substrates. This serine protease, which has previously been isolated as cDNA from a catfish natural killer-like cell line showed a preference for Ala in the P1 position of the substrate, and for multiple basic amino acids N-terminally of the cleavage site. A closely related zebrafish serine protease (zebrafish esterase-like) showed a very similar cleavage specificity, indicating an evolutionary conservation of this protease specificity among various fish species. Two catfish serine proteases, originating from NK-like cells, have now been isolated and characterized. One of them is highly specific met-ase with similar characteristics as the mammalian granzyme M. This enzyme may be involved in the induction of apoptosis in virus-infected cells, with a potential target in (catfish) caspase 6. In contrast to catfish granzyme-like I, the second enzyme analyzed here does not seem to have a direct counterpart in mammalian NK cells, and its role in the immune function of catfish NK cells is, therefore, still not known. However, this enzyme seems to be able to cleave a number of cytoskeletal proteins, indicating a separate strategy to induce apoptosis in target cells. Both of these enzymes are very interesting targets for further studies of their roles in catfish immunity, as enzymes with similar specificities have also been identified in zebrafish.

Nanostructured anti-reflection coating for absorption enhancement in perovskite silicon tandem solar cells

Perovskite-silicon tandem solar cells have captured the attention of the solar cell research community due to the advantages of perovskites, such as, an easy fabrication process using sol-gel methods and silicon bottom cells that can be fabricated using well-established fabrication techniques. The present study discusses the design, optimization, and numerical analysis related to the role of nanostructured anti-reflection coating design for perovskite (MAPbI3) silicon tandem solar cells. In the design, the top cell is taken as MAPbI3 and the bottom cell is C-silicon. The anti-reflection coating is designed with SiO2 nanoparticles embedded in ITO. These nanostructured top anti-reflection coating results are compared with its planar top cell counterpart. SiO2 nanoparticle diameter and interparticle separation are optimized to get maximum absorption in the top cell. Upon optimization, it was found that a design having SiO2 nanoparticles with a diameter of 60 nm and no interparticle separation showed the most reduction in reflection, which in turn led to an increase in absorption in the top cell. The proposed structure enhances current density by 8.3% over the planar cell. This top cell current is matched to the bottom silicon thickness. These findings were validated using Mie scattering and the Bruggmann effective medium approximation.

The crosstalk between ILC3s and adaptive immunity in diseases.

RORγt+ group 3 innate lymphoid cells (ILC3s), the innate counterpart of Th17 cells, are enriched in the mucosal area and lymphoid tissues. ILC3s interact with a variety of cells through their effector molecules and play an important role in the host defense against a spectrum of infections. Recent studies suggest that the extensive crosstalk between ILC3s and adaptive immune cells, especially T cells, is essential for maintaining tissue homeostasis. Here we discuss recent advances in the crosstalk between ILC3s and adaptive immune responses in multiple tissues and diseases. Understanding how ILC3s engage with adaptive immune cells will enhance our comprehension of diseases and facilitate the identification of novel therapeutic targets.

Modeling Contact Resistance and Water Transport within a Cathode Liquid-Fed Proton Exchange Membrane Electrolyzer

Conventional proton-exchange membrane (PEM) water electrolyzers use thicker membranes (>175 μm) than their PEM fuel cell counterparts (<25 μm), which reduces hydrogen crossover but also reduces electrolyzer efficiency due to the increased resistance. Reduction of hydrogen crossover is critical in conventional systems to avoid buildup of hydrogen in the anode above the lower flammability limit. New concepts for operating PEM water electrolyzers are emerging, such as the patented concept involving liquid water supply at the cathode while operating the anode with air, which reduces the safety concern related to hydrogen crossover using thin membranes. Experimental work has demonstrated the viability of this approach, but open questions remain regarding the interplay between water transport, water consumption, and cell performance, as well as identifying the components and material properties that enable high performance. In this work, a physics-based computational model of a cathode-fed PEM water electrolyzer was developed. The model highlights the importance of limiting contact resistance and explores the effect of cell compression on non-uniformity of current distributions. Sensitivity studies found that membranes up to 50 μm thick can be used without significant water transport limitations.

Abstract A020: Reversing triple negative breast cancer stem cells by disrupting the Notch signaling pathway via BCL6 targeting

Abstract Triple negative breast cancer (TNBC) is a heterogeneous molecular breast cancer (BC) subtype that still has a very poor prognosis. Genetic, molecular and clinical tumor heterogeneity represent major pathobiological factors involved in poor clinical outcome. The sequence specific transcriptional repressor and proto-oncogene B cell lymphoma protein 6 (BCL6) has emerged as a suitable therapeutic target as it plays broad and critical roles in many hematological and solid cancers. BCL6 exerts control over cellular proliferation and survival by binding to specific responsive elements located within the regulatory regions of target genes and recruiting of chromatin modifying co-repressor complexes such as silencing mediator for retinoid and thyroid receptors (SMRT), BCL6 corepressor (BCoR) and histone deacetylases (HDACs). Moreover, in hematological malignancies, BCL6 transcriptional activity is also associated to the recruitment of the chromatin remodeling factor, Enhancer of Zeste 2 (EZH2). The role of BCL6 in BC is still not well defined; however emerging studies report its involvement in tumorigenesis, disease progression and poor survival. We recently showed that BCL6 is expressed at significantly higher levels in TNBC mammospheres, 3D multicellular structures enriched in cancer stem cells (CSCs), than in their differentiated bulk cell counterparts. Additionally, we found that the functional role of BCL6 is crucial for mammosphere formation, as its loss of function decreased this process. Intriguingly, we uncovered a functional interplay occurring between BCL6 and EZH2, which triggers the BCL6/Notch stemness signaling axis by inhibiting Numb transcription in both SUM149 and SUM159 TNBC cell lines. We exploited the molecular mechanisms responsible for this specific interplay, taking advantage of two molecules, FX1, a small molecule inhibitor able to block BCL6 recruiting activity and EED226, a potent Polycomb Repressive Complex 2 (PRC2) inhibitor. In order to define the role of both BCL6, as epigenetic regulator, and NUMB, as inhibitor of NOTCH stemness pathway in TNBC, SUM149 and SUM159 cell lines were stably transduced with lentiviral shRNA to derive stable NUMB-silenced matched cells. The expression of Notch downstream target genes, such as HES1/2 and HEY1/2, was assessed by qRT PCR analysis in the engineered TNBC cell models and resulted only slightly increased following NUMB downmodulation. Upon FX1 or EED226 administration, a strongest effect was observed, indicating that NUMB is not the only player in regulating the Notch pathway. Further experiments are currently ongoing aiming at identify additional players and molecular pathway(s). Collectively, our preliminary results provide additional preclinical evidences on the role exerted by BCL6 as epigenetic regulator of TNBC and underlie its interplay with EZH2thus paving the way for a novel potential therapeutic strategy targeting BCL6/Notch signaling axis. Citation Format: Francesca De Santis, Francesca Putti, Giorgia Bravin, Carla Portulano, Federica Zonca, Lorenzo Castagnoli, Michele Magni, Filippo de Braud, Serenella M Pupa, Massimo Di Nicola. Reversing triple negative breast cancer stem cells by disrupting the Notch signaling pathway via BCL6 targeting [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A020.