Building a Better Defense: Expanding and Improving Natural Killer Cells for Adoptive Cell Therapy.

Natural killer (NK) cells have gained attention as a promising adoptive cell therapy platform for their potential to improve cancer treatments. NK cells offer distinct advantages over T-cells, including major histocompatibility complex class I (MHC-I)-independent tumor recognition and low risk of toxicity, even in an allogeneic setting. Despite this tremendous potential, challenges persist, such as limited in vivo persistence, reduced tumor infiltration, and low absolute NK cell numbers. This review outlines several strategies aiming to overcome these challenges. The developed strategies include optimizing NK cell expansion methods and improving NK cell antitumor responses by cytokine stimulation and genetic manipulations. Using K562 cells expressing membrane IL-15 or IL-21 with or without additional activating ligands like 4-1BBL allows "massive" NK cell expansion and makes multiple cell dosing and "off-the-shelf" efforts feasible. Further improvements in NK cell function can be reached by inducing memory-like NK cells, developing chimeric antigen receptor (CAR)-NK cells, or isolating NK-cell-based tumor-infiltrating lymphocytes (TILs). Memory-like NK cells demonstrate higher in vivo persistence and cytotoxicity, with early clinical trials demonstrating safety and promising efficacy. Recent trials using CAR-NK cells have also demonstrated a lack of any major toxicity, including cytokine release syndrome, and, yet, promising clinical activity. Recent data support that the presence of TIL-NK cells is associated with improved overall patient survival in different types of solid tumors such as head and neck, colorectal, breast, and gastric carcinomas, among the most significant. In conclusion, this review presents insights into the diverse strategies available for NK cell expansion, including the roles played by various cytokines, feeder cells, and culture material in influencing the activation phenotype, telomere length, and cytotoxic potential of expanded NK cells. Notably, genetically modified K562 cells have demonstrated significant efficacy in promoting NK cell expansion. Furthermore, culturing NK cells with IL-2 and IL-15 has been shown to improve expansion rates, while the presence of IL-12 and IL-21 has been linked to enhanced cytotoxic function. Overall, this review provides an overview of NK cell expansion methodologies, highlighting the current landscape of clinical trials and the key advancements to enhance NK-cell-based adoptive cell therapy.

Genetic Manipulation Approaches to Enhance the Clinical Application of NK Cell-Based Immunotherapy.

Natural killer (NK) cells are a subset of cytotoxic lymphocytes within the innate immune system. While they are naturally cytotoxic, genetic modifications can enhance their tumor-targeting capability, cytotoxicity, persistence, tumor infiltration, and prevent exhaustion. These improvements hold the potential to make NK-cell-based immunotherapies more effective in clinical applications. Currently, several viral and non-viral technologies are used to genetically modify NK cells. For nucleic acid delivery, non-viral methods such as electroporation, lipid nanoparticles, lipofection, and DNA transposons have gained popularity in recent years. On the other hand, viral methods including lentivirus, gamma retrovirus, and adeno-associated virus, remain widely used for gene delivery. Furthermore, gene editing techniques such as clustered regularly interspaced short-palindromic repeats-based, zinc finger nucleases, and transcription activator-like effector nucleases are the pivotal methodologies in this field. This review aims to provide a comprehensive overview of chimeric antigen receptor (CAR) arming strategies and discuss key gene editing techniques. These approaches collectively aim to enhance NK cell/NK cell CAR-based immunotherapies for clinical translation.

B Cells in the Gastrointestinal Tumor Microenvironment with a Focus on Pancreatic Cancer: Opportunities for Precision Medicine?

We review state-of-the-art in translational and clinical studies focusing on the tumor microenvironment (TME) with a focus on tumor-infiltrating B cells (TIBs). The TME is a dynamic matrix of mutations, immune-regulatory networks, and distinct cell-to-cell interactions which collectively impact on disease progress. We discuss relevant findings concerning B cells in pancreatic cancer, the concepts of "bystander" B cells, the role of antigen-specific B cells contributing to augmenting anticancer-directed immune responses, the role of B cells as prognostic markers for response to checkpoint inhibitors (ICBs), and the potential use in adoptive cell tumor-infiltrating lymphocyte (TIL) products.

Trained Immunity for Personalized Cancer Immunotherapy: Current Knowledge and Future Opportunities.

Memory formation, guided by microbial ligands, has been reported for innate immune cells. Epigenetic imprinting plays an important role herein, involving histone modification after pathogen-/danger-associated molecular patterns (PAMPs/DAMPs) recognition by pattern recognition receptors (PRRs). Such "trained immunity" affects not only the nominal target pathogen, yet also non-related targets that may be encountered later in life. The concept of trained innate immunity warrants further exploration in cancer and how these insights can be implemented in immunotherapeutic approaches. In this review, we discuss our current understanding of innate immune memory and we reference new findings in this field, highlighting the observations of trained immunity in monocytic and natural killer cells. We also provide a brief overview of trained immunity in non-immune cells, such as stromal cells and fibroblasts. Finally, we present possible strategies based on trained innate immunity that may help to devise host-directed immunotherapies focusing on cancer, with possible extension to infectious diseases.

Metodologias de intervenção do terapeuta ocupacional em contexto escolar com crianças com Necessidades Educativas Especiais em Portugal / Occupational therapists intervention methodologies in schools with children with Special Educational Needs in Portugal

Introdução: Os alunos com Necessidades Educativas Especiais devem ser inseridos em turmas regulares, recebendo todos os serviços educativos adequados que possibilitem o desenvolvimento de competências e capacidades dos mesmos. O sucesso da inclusão escolar depende da parceria entre os vários profissionais a intervir em contexto escolar. A importância do terapeuta ocupacional inserido numa equipe em contexto escolar já é reconhecida e valorizada pela comunidade, e é alvo de muitos estudos. Objetivo: Identificar as metodologias de intervenção utilizadas pelos terapeutas ocupacionais em Portugal, em contexto escolar, com crianças com Necessidades Educativas Especiais, de forma a aferir as metodologias mais comuns, bem como os aspectos relevantes que fazem parte do processo terapêutico. Método: Trata-se de uma investigação de caráter não experimental com base descritiva e transversal. Inicialmente, foi elaborado um questionário semiestruturado, de natureza quantitativa, que foi ministrado aos terapeutas ocupacionais que trabalham ou tenham trabalhado havia menos de três anos em contexto escolar. Resultados: A amostra é composta por 40 terapeutas ocupacionais, 37 são do sexo feminino e três são do sexo masculino. A maioria dos indivíduos (77,5%) trabalha atualmente em contexto escolar. Os terapeutas ocupacionais intervêm principalmente com crianças, cujas idades estão compreendidas entre os 6 e os 18 anos. Conclusão: Verifica-se que as metodologias mais utilizadas pelos terapeutas ocupacionais são a atividade lúdica/brincar terapêutico e o treino de atividades da vida diária. A equitação com fins terapêuticos e a hipoterapia representam as metodologias menos utilizadas em contexto escolar, em que cada uma foi selecionada por 10% dos terapeutas.

Introduction: students with special educational needs should be inserted in regular classes, receiving all appropriate educational services allowing the development of skills and capabilities. The success of the school inclusion depends on the partnership between the various professionals in the school context. The insertion and occupational therapist's contribution in the area of education is the subject of many studies. The importance of a professional team in school context is already recognized and valued by the community. Objectives: the aim is to identify the intervention methodologies used by occupational therapists in school with children with special educational needs. In order to determine the most common methodologies as well as relevant aspects of the therapeutic process. Method: this is a non-experimental research with descriptive and cross-sectional basis. A semi-structured questionnaire was prepared initially, quantitative in nature that was taught to occupational therapists that work or have worked for less than 3 years in school context. Results: the sample is composed of 40 occupational therapists, 37 are female and 3 are male. The majority of individuals (77.5%) currently works in school context. The occupational therapists involved mainly with children, aged between 6 and 18 years. Conclusion: the methodologies used by the occupational therapists are playful activity/play therapy and the training activities of daily living. Riding for therapeutic purposes and the hippotherapy represent less widely used methodologies in school context, in that each was selected by 10% of therapists.