Targeted delivery of potent cytotoxic drugs to cancer cells minimizes systemic toxicity and several side effects. NHC*−Au−Cl has already been proven to be a potent anticancer agent. In this study, we ...explore a strategy based on chemoselective cysteine conjugation of NHC*−Au−Cl to albumin and trastuzumab (Thiomab LC‐V205C) to potentiate drug‐ligand ratio, pharmacokinetics, as well as drug efficacy and safety. This strategy is a step forward towards the use of gold‐based anticancer agents as targeted therapies.
Golden therapy: Synthesis of NHC*−Au−S‐protein bioconjugates to increase stability, modulate pharmacokinetics, and ultimately enhance efficacy and safety of gold‐based anticancer drugs. The findings reported represent a step forward towards the use of gold‐based anticancer drugs as targeted therapies.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
The impact of nutritional status during fetal life on the overall health of adults has been recognized; however, dietary effects on the developing immune system are largely unknown. Development of ...secondary lymphoid organs occurs during embryogenesis and is considered to be developmentally programmed. Secondary lymphoid organ formation depends on a subset of type 3 innate lymphoid cells (ILC3) named lymphoid tissue inducer (LTi) cells. Here we show that mouse fetal ILC3s are controlled by cell-autonomous retinoic acid (RA) signalling in utero, which pre-sets the immune fitness in adulthood. We found that embryonic lymphoid organs contain ILC progenitors that differentiate locally into mature LTi cells. Local LTi cell differentiation was controlled by maternal retinoid intake and fetal RA signalling acting in a haematopoietic cell-autonomous manner. RA controlled LTi cell maturation upstream of the transcription factor RORγt. Accordingly, enforced expression of Rorgt restored maturation of LTi cells with impaired RA signalling, whereas RA receptors directly regulated the Rorgt locus. Finally, we established that maternal levels of dietary retinoids control the size of secondary lymphoid organs and the efficiency of immune responses in the adult offspring. Our results reveal a molecular link between maternal nutrients and the formation of immune structures required for resistance to infection in the offspring.
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IJS, KISLJ, NUK, UL, UM, UPUK
We describe a de novo computational approach for designing proteins that recapitulate the binding sites of natural cytokines, but are otherwise unrelated in topology or amino acid sequence. We use ...this strategy to design mimics of the central immune cytokine interleukin-2 (IL-2) that bind to the IL-2 receptor βγ
heterodimer (IL-2Rβγ
) but have no binding site for IL-2Rα (also called CD25) or IL-15Rα (also known as CD215). The designs are hyper-stable, bind human and mouse IL-2Rβγ
with higher affinity than the natural cytokines, and elicit downstream cell signalling independently of IL-2Rα and IL-15Rα. Crystal structures of the optimized design neoleukin-2/15 (Neo-2/15), both alone and in complex with IL-2Rβγ
, are very similar to the designed model. Neo-2/15 has superior therapeutic activity to IL-2 in mouse models of melanoma and colon cancer, with reduced toxicity and undetectable immunogenicity. Our strategy for building hyper-stable de novo mimetics could be applied generally to signalling proteins, enabling the creation of superior therapeutic candidates.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
T helper (Th) cells are critical players in the modulation of immune response outcomes. Activation of Th cells gives rise to various subsets of effector cells that are controlled via specialised ...regulatory T cells or through self‐regulation via production of IL‐10. However, the environmental factors that regulate IL‐10 production by Th cells remain poorly understood. Here, we show that the neurotrophic factor receptor rearranged during transfection (RET) downregulates IL‐10 production by Th cells from C57BL/6 mice. We found that effector Th cells express RET and that RET's neurotrophic factor partners are mainly produced by LN stromal cells, allowing context‐dependent Th‐cell regulation. Despite being dispensable for Th‐cell homeostasis, RET controls IL‐10 production in Th2 cells: RET‐deficient Th cells exhibited increased IL‐10 production, while triggering of Th1/2 cells with neurotrophic factors, namely glial‐derived neurotrophic factor and neurturin, decreased the expression of IL‐10. In agreement, the important IL‐10 transcription factor Maf was upregulated in RET‐deficient Th2 cells and down‐regulated upon RET signalling activation by glial‐derived neurotrophic factor family ligands. Thus, our study uncovers neurotrophic factors as novel regulators of Th‐cell function, revealing that Th cells and neurons can be regulated by similar signals in tissue‐specific responses.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Abstract
The emergence of novel variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) underscores the need to investigate alternative approaches to prevent infection and treat ...patients with coronavirus disease 2019. Here, we report the preclinical efficacy of NL-CVX1, a de novo decoy that blocks virus entry into cells by binding with nanomolar affinity and high specificity to the receptor-binding domain of the SARS-CoV-2 spike protein. Using a transgenic mouse model of SARS-CoV-2 infection, we showed that a single prophylactic intranasal dose of NL-CVX1 conferred complete protection from severe disease following SARS-CoV-2 infection. Multiple therapeutic administrations of NL-CVX1 also protected mice from succumbing to infection. Finally, we showed that infected mice treated with NL-CVX1 developed both anti-SARS-CoV-2 antibodies and memory T cells and were protected against reinfection a month after treatment. Overall, these observations suggest NL-CVX1 is a promising therapeutic candidate for preventing and treating severe SARS-CoV-2 infections.
We show that NL-CVX1, a de novo human angiotensin-converting enzyme 2 protein decoy that blocks viral cell invasion, prevents severe disease after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in mice and may constitute a potent anti–SARS-CoV-2 treatment.
Bruton’s tyrosine kinase (BTK) is a member of the TEC-family kinases and crucial for the proliferation and differentiation of B-cells. We evaluated the therapeutic potential of a covalent inhibitor ...(JS25) with nanomolar potency against BTK and with a more desirable selectivity and inhibitory profile compared to the FDA-approved BTK inhibitors ibrutinib and acalabrutinib. Structural prediction of the BTK/JS25 complex revealed sequestration of Tyr551 that leads to BTK’s inactivation. JS25 also inhibited the proliferation of myeloid and lymphoid B-cell cancer cell lines. Its therapeutic potential was further tested against ibrutinib in preclinical models of B-cell cancers. JS25 treatment induced a more pronounced cell death in a murine xenograft model of Burkitt’s lymphoma, causing a 30–40% reduction of the subcutaneous tumor and an overall reduction in the percentage of metastasis and secondary tumor formation. In a patient model of diffuse large B-cell lymphoma, the drug response of JS25 was higher than that of ibrutinib, leading to a 64% “on-target” efficacy. Finally, in zebrafish patient-derived xenografts of chronic lymphocytic leukemia, JS25 was faster and more effective in decreasing tumor burden, producing superior therapeutic effects compared to ibrutinib. We expect JS25 to become therapeutically relevant as a BTK inhibitor and to find applications in the treatment of hematological cancers and other pathologies with unmet clinical treatment.
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IJS, KILJ, NUK, PNG, UL, UM, UPUK
Changes in diet and microbiota have determining effects on the function of the mucosal immune system. For example, the active metabolite of vitamin A, retinoic acid (RA), has been described to ...maintain homeostasis in the intestine by its influence on both lymphocytes and myeloid cells. Additionally, innate lymphoid cells (ILCs), important producers of cytokines necessary for intestinal homeostasis, are also influenced by vitamin A in the small intestines. In this study, we show a reduction of both NCR(-) and NCR(+) ILC3 subsets in the small intestine of mice raised on a vitamin A-deficient diet. Additionally, the percentages of IL-22-producing ILCs were reduced in the absence of dietary vitamin A. Conversely, mice receiving additional RA had a specific increase in the NCR(-) ILC3 subset, which contains the lymphoid tissue inducer cells. The dependence of lymphoid tissue inducer cells on vitamin A was furthermore illustrated by impaired development of enteric lymphoid tissues in vitamin A-deficient mice. These effects were a direct consequence of ILC-intrinsic RA signaling, because retinoic acid-related orphan receptor γt-Cre × RARα-DN mice had reduced numbers of NCR(-) and NCR(+) ILC3 subsets within the small intestine. However, lymphoid tissue inducer cells were not affected in these mice nor was the formation of enteric lymphoid tissue, demonstrating that the onset of RA signaling might take place before retinoic acid-related orphan receptor γt is expressed on lymphoid tissue inducer cells. Taken together, our data show an important role for vitamin A in controlling innate lymphoid cells and, consequently, postnatal formed lymphoid tissues within the small intestines.
A mucosa intestinal abriga uma vasta comunidade de mhomo icroorganismos que contribuem para o normal funcionamento dos organismos superiores. Encontra-se dotada de um sistema imunitário que se ...compartimentaliza em diferentes estruturas linfóides que permitem a interacção com o meio exterior. O sistema imune intestinal contribui para a homeostase intestinal, tolerando os organismos comensais e definindo respostas inflamatórias contra patogénios. Num contexto homeostático, o estabelecimento de mecanismos que regulam o desenvolvimento de respostas imunes após interacção com um organismo ou antigénio desconhecido tornam-se indispensáveis. A influência de nutrientes no estabelecimento e regulação destas respostas tem vindo a ser demostrada nos últimos anos. Contudo, pouco ainda é conhecido sobre os mecanismos moleculares subjacentes à interacção entre nutrientes e as células imunes.A deficiência em vitamina A afecta aproximadamente 250 milhões de pessoas em todo o mundo, sendo responsável por um grave aumento de morbidez a infecções gastrointestinais e respiratórias. O ácido retinóico (RA) é um metabolito da vitamina A que desempenha importantes funções no desenvolvimento do sistema imunitário e na regulação das respostas imunes. Uma resposta imunitária adequada requer a existência de mecanismos moleculares que permitam o rápido recrutamento de células imunes para combater o alastramento de uma infecção. Entre estes mecanismos inclui-se a aquisição da expressão de moléculas designadas por marcadores de residência que permitem o reconhecimento de sinais quimiotácticos e a retenção das células num determinado tecido.O receptor CCR9 e as integrinas α4β7 são os marcadores de residência mais importantes para o estabelecimento de células T na mucosa intestinal. Estudos anteriores relatam que o processo de aquisição da expressão de CCR9 e α4β7 é promovido por células dendríticas (DCs) CD103pos provenientes do intestino que metabolizam a vitamina A e fornecem RA a células T na periferia. Nas células T, o RA interage com receptores do ácido retinóico (RAR) desencadeando vias de sinalização que induzem a expressão de CCR9 e de α4β7. Os linfócitos intra-epiteliais (IELs) residem no epitélio e constituem a primeira linha de combate do sistema imunitário nas mucosas. Estes linfócitos caratecterizam-se em duas classes distintas: IELs não convencionais (ou naturais) - linfócitos T γδ e T αβ que expressam o homodímero CD8αα, e células T γδ que não expressam CD8 - e convencionais (ou induzidas) – compostas pelas células T CD4pos e CD8posclássicas.Apesar da função do RA no recrutamento e estabelecimento de células T na lamina própria intestinal já ter sido demonstrada, pouco é sabido sobre o seu papel na função e migração dos IELs, como também sobre a própria dinâmica migracional destas células. Mais ainda, estas experiências foram realizadas através de dietas pobres em vitamina A, depletando o RA de forma promíscua. Por esta razão propusémo-nos a estudar o papel do RA na função e estabelecimento de células T no epitélio intestinal. Para tal, usámos ratinhos que possuem uma forma do RARα truncada após o aminoácido 403 (RARaT403) inserida no gene ubiquamente expresso ROSA-26, com um codão stop flanqueado por sequências loxP (ratinhos T403). O RARaT403 é incapaz de iniciar a cascata de sinalização a jusante do RARα e possui dominância negativa para com todas as isoformas do RAR. De forma a bloquear a sinalização do RA em células T, cruzámos estes ratinhos com outros que possuem um recombinase Cre, que reconhece sequências loxP, sob controlo do promotor do gene CD2 humano (ratinhos CD2cre T403).
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