Auswahl der wissenschaftlichen Literatur zum Thema „Car-Vst“

Abstract Donor lymphocyte infusions (DLI) following hematopoietic stem cell transplantation may reduce or control opportunistic infections and leukemia/lymphoma relapse, but the associated graft versus host disease (GvHD) limits the clinical success of this procedure. Since T cell immunotherapy may be a safer alternative to DLI we have now used a single T cell platform that mediates both antileukemic and antiviral activity. Autologous T cells modified to express CD19-specific chimeric antigen receptors (CD19.CAR) have had clinical activity against CD19-expressing malignancies, but it is unknown if similarly modified allogeneic T cells will be equally effective. Allogeneic virus specific T cells (VSTs) directed to cytomegalovirus (CMV), adenovirus (Adv), and Epstein Barr virus (EBV) have been shown to be safe and effective in preventing and treating life-threatening viral infections post HSCT. Therefore, we sought to determine whether allogeneic VSTs could be engineered to express CD19.CAR and would retain the safety and effectiveness of unmodified VSTs whilst gaining anti-tumor activity. VSTs were expanded ex vivo using antigen presenting cells engineered to express adenovirus and cytomegalovirus (using an Ad5f35 adenoviral vector expressing the CMV pp65 gene), and Epstein Barr virus (using EBV-infected lymphoblastoid cell lines) antigens. After 3 stimulations, the VST’s were modified to express CD19.CAR.28ζ using a retroviral vector encoding the CAR-CD19 receptor coupled to the CD28 co-stimulatory molecule and the T cell receptor zeta (ζ) chain. Nine CD19.CAR-modified virus specific T cell (CD19.CAR-VSTs) products were generated for infusion. All VST lines recognized at least one viral antigen as determined by Elispot or chromium release assays and 20% to 48% of cells expressed the CD19.CAR. All lines killed CD19-expressing cells in vitro. We treated nine patients with these CD19.CAR-VSTs, 3 months to 13 years after HSCT. Six patients received CD19.CAR-VSTs for relapsed disease and 3 patients received the T cells as adjuvant therapy to prevent viral infection and relapse after HSCT. Safety. There were no infusion-related toxicities. One patient presented with gastrointestinal symptoms following infusion subsequently determined to be unrelated to the T cells. Persistence. VSTs persisted a median of 8 weeks in the peripheral blood and up to 9 weeks at disease sites. In three patients (#1, #3 and #5), CD19.CAR signals were detectable in the bone marrow or the lymph nodes (44.8, 25.85, and 32 copies/1000 ng DNA) even when no signal was measurable in peripheral blood, indicating preferential accumulation of the infused T cells at the disease site. Anti-Tumor Activity. During the period of CD19.CAR-VST persistence, objective anti-tumor activity was evident in 2/6 patients with relapsed disease (patient # 1 had detectable blasts in the peripheral blood which disappeared within 1-2 weeks following infusion, patient # 2 had 16% circulating CLL cells which decreased within 2 weeks of T cell infusion) but disease recurred after 3 and 2 months, respectively. The two patients who received cells while in remission remain disease-free >3 and >9 months later. Anti-Viral Activity. In two patients with EBV reactivation, donor CD19.CAR-VSTs expanded concomitant with an increase in virus-specific T cell responses, and decreased viral load. A third patient had a rise in adenovirus specific VSTs during an episode of adenovirus associated diarrhea. Although the infection was controlled, there was no concomitant rise in CD19-CAR expressing T cells in this patient. No other patient had viral disease. In conclusion, allogeneic CD19.CAR-VSTs administered after allogeneic HSCT are safe and can exert both anti-tumor and anti-viral activity in the absence of GvHD. Earlier administration of CD19.CAR-VSTs after HSCT, when the host is lymphodepleted and the incidence of viral infection is higher, may allow these cells to better capture the potential advantages of native TCR stimulation (and associated co-stimulation) for expansion and persistence, and thereby produce a higher frequency of sustained tumor responses. Alternatively, intentional stimulation of the native TCRs by viral vaccines may produce equal benefit, with greater predictability. Disclosures: Savoldo: Celgene: Patents & Royalties, Research Funding. Heslop:Celgene: Patents & Royalties, Research Funding; Cell Medica: Patents & Royalties. Rooney:Cell Medica: Membership on an entity’s Board of Directors or advisory committees, Patents & Royalties, Research Funding; Celgene: Patents & Royalties, Research Funding. Brenner:Celgene: Patents & Royalties, Research Funding. Dotti:Celgene: Patents & Royalties, Research Funding.

TPS2679 Background: Urgent therapies are needed for patients with refractory Human Epidermal Growth Factor Receptor 2 (HER2) positive solid malignancies. Our group has previously shown potent antitumor effect of a binary oncolytic/helper-dependent adenovirus (CAdVEC) with dual expression of IL-12 and PD-L1 blocker. Initial preclinical and clinical studies with direct tumor injection of CAdVEC has shown it to be safe and result in increased infiltration of CD8 T cells into the tumor microenvironment, with antitumor effect on locoregional and distant metastatic sites.1 Based on these results, we describe an ongoing study with addition of HER2-specific autologous chimeric antigen receptor (CAR)-T cell therapy to CAdVEC. Methods: This is a single arm, dose escalation phase I clinical trial guided by Bayesian Optimal Interval Design (BOIN) for patients with advanced HER2 positive solid tumors. Patients who are deemed unsuitable for curative treatments and progressed after at least one standard first line therapy are eligible. HER2 positivity is determined by IHC and defined as ≥2+. Patients are required to have at least one tumor site appropriate for intratumor injection and radiographically measurable disease per RECIST v1.1. Patients with autoimmune disease requiring systemic corticosteroids greater than 10mg/day and those with active/untreated CNS metastasis are excluded. Patients will receive increasing doses of CAdVEC intratumor injection alone (first two dose levels) or in combination with HER2 specific autologous CAR-T cells (dose levels 3-7). A total of 45 patients are planned. The primary endpoint of the study is to evaluate safety and maximum tolerated dose as assessed by incidence of dose limiting toxicities (DLT) of CAdVEC intratumor injection in combination with HER2-specific autologous CAR T cells. Secondary endpoints include anti-tumor activity of the combination measured by overall response rate (ORR), disease control rate (DCR), median progression free survival (PFS) and median overall survival (OS). Exploratory endpoints include assessing the immunogenicity of combination therapy by determining the impact of treatment on cellular and humoral immunity, as well as assessing long-term persistence and functional status of HER2 CAR T-cells. Correlative analysis of levels of adenovirus antibodies with clinical outcomes and immunological findings are also planned. This study is currently enrolling. 1. Wang D, Porter CE, Lim B, et al: Science Advances 9, 2023. Clinical trial information: NCT03740256 .

Adoptive cell therapy (ACT) has transformed the treatment landscape for cancer and infectious disease through the investigational use of chimeric antigen receptor T-cells (CAR-Ts), tumour-infiltrating lymphocytes (TILs) and viral-specific T-cells (VSTs). Whilst these represent breakthrough treatments, there are subsets of patients who fail to respond to autologous ACT products. This is frequently due to impaired patient T-cell function or “fitness” as a consequence of prior treatments and age, and can be exacerbated by complex manufacturing protocols. Further, the manufacture of autologous, patient-specific products is time-consuming, expensive and non-standardised. Induced pluripotent stem cells (iPSCs) as an allogeneic alternative to patient-specific products can potentially overcome the issues outlined above. iPSC technology provides an unlimited source of rejuvenated iPSC-derived T-cells (T-iPSCs) or natural killer (NK) cells (NK-iPSCs), and in the context of the growing field of allogeneic ACT, iPSCs have enormous potential as a platform for generating off-the-shelf, standardised, “fit” therapeutics for patients. In this review, we evaluate current and future applications of iPSC technology in the CAR-T/NK, TIL and VST space. We discuss current and next-generation iPSC manufacturing protocols, and report on current iPSC-based adoptive therapy clinical trials to elucidate the potential of this technology as the future of ACT.

Les portes semblent se refermer sur bon nombre d’espaces cliniques. C’est une oppression insidieuse, car elle infiltre notre propre quotidienneté, inonde nos schémas de penser, nos formes d’existence. D’aucuns l’ont nommée une « dictature du On ». Dès lors, comment résister à cette chasse des mouvements d’émancipation, sans s’isoler dans le passage à l’acte ? Il semblerait qu’une voie intermédiaire se dessine, celle d’une désobéissance clinique.

Georges Canguilhem (1904-1995) fut philosophe, médecin et résistant. Célèbre notamment pour son ouvrage Le normal et le pathologique , rédigé pendant la guerre, il propose une défense de la clinique fondée sur le respect de la normativité propre à chaque individu, éclairée par la physiologie. Moins connus car restés longtemps inédits, ses travaux sur la psychologie des années 1930 tissent un lien entre maladie mentale et liberté dont découle une forme de soin psychologique vigilant à soutenir les efforts thérapeutiques du patient.

Le soutien des projets de médiation, qu’ils soient artistiques, sportifs, artisanaux et/ou professionnels, est incontournable dans le travail d’accompagnement des personnes en situation de précarité, que cet accompagnement soit effectué par des professionnels (du travail social), ou par des citoyen·nes (sous-entendu « bénévoles »). Car il se joue à cet endroit l’épanouissement de la personne, son intégration à des réseaux de personnes et la revalorisation de son image dans la complexité des représentations qui la concernent. Merci à Rodney P. de nous en faire prendre conscience à travers son témoignage.

Основным источником загрязнения поверхностного стока в крупных городах является автотранспорт, поэтому стоки с автомобильных дорог и автопарковок необходимо подвергать глубокой очистке, включающей несколько ступеней. В рамках решения данной проблемы была проведена работа по оценке эксплуатации двухкаскадной установки (на основе последовательно работающих фильтров ФОПС®-С и ФОПС®-МУ) при очистке поверхностного стока, поступающего с территории автопарковки при различных сезонных изменениях. В результате тестовых испытаний доказано, что на протяжении 21 месяца установка обеспечивала эффективность очистки поверхностного стока от взвешенных веществ 98–99% и от нефтепродуктов 94–97%. После эксплуатации в течение трех лет, включая зимние периоды с отрицательными температурами до менее –20 °С, установка обеспечивала требуемую эффективность очистки по контролируемым параметрам. Периодически производилась очистка фильтра ФОПС®-С – выгрузка взвешенных веществ и нефтепродуктов. Moto transport has been the main source of pollution of surface runoff in large cities; therefore, runoff from roads and car parking spaces must be subjected to enhanced treatment including several stages. As part of solving this problem, works were carried out to estimate the operation of a two-stage plant (based on sequentially operating FOPS®-S and FOPS®-MU filters) in the process of surface runoff treatment from the car parking spaces during various seasonal changes. As a result of test experiments, it was found that for 21 months the plant provided for 98–99% efficiency of removing suspended solids and 94–97% efficiency of removing oil products from the surface runoff. After three years operation including winter periods with below-zero temperatures to less than –20 °C, the plant provided for the required treatment level according to the monitored parameters. Occasionally, the FOPS®-S filter was cleaned – removing suspended solids and oil products.

La thérapie CAR-T a transformé le traitement du cancer en modifiant les lymphocytes T pour cibler spécifiquement les antigènes tumoraux. Bien que cette approche ait montré un succès remarquable dans les hémopathies malignes à cellules B, le processus reste coûteux et long, car il nécessite la collecte et la modification des cellules du patient, ce qui peut retarder le traitement. De plus, certains patients, en raison de traitements antérieurs ou de maladies avancées, ne disposent pas de cellules viables, limitant l'accès à cette thérapie.Les cellules CAR-T allogéniques provenant de donneurs offrent une solution plus rapide et évolutive, réduisant le temps de production et les coûts. Cependant, elles présentent des risques, notamment la maladie du greffon contre l'hôte (GvHD), où les cellules du donneur attaquent les tissus du patient. Notre étude a exploré une approche innovante, combinant la technologie CAR avec des lymphocytes T spécifiques aux virus (Virus Specific T cells, VST), connus pour leurs propriétés antivirales et antitumorales, afin de générer des CAR-VST. Ces CAR-VST à double spécificité représentent une alternative prometteuse, particulièrement pour les patients à risque de rechute tumorale ou de réactivation virale.Dans notre étude, nous avons généré des CAR-T et des CAR-VST à partir des mêmes donneurs, obtenant respectivement 40,28%±9,30% et 35,96%±11,40% d'expression de CD19.CAR au jour 7 (N=3). Les CAR-VST ont montré in vitro une clairance tumorale similaire à celle des CAR-T, avec 74,13%±22,06% de lyse des cellules CD19+. Dans un modèle murin, un contrôle de la croissance tumorale ainsi qu'une amélioration de la survie similaires ont été observés dans les deux groupes. De plus, les CAR-VST ont conservé leur activité antivirale, lysant 62,32%±13,84% des cellules chargées en peptides viraux. Concernant l'alloréactivité, les CAR-VST ont montré une prolifération CD3+ inférieure (28,27%±21,64%) par rapport aux CAR-T (88,3%±24,48%, p=0,0285, N=4), suggérant un risque réduit de GvHD.En collaboration avec l'Université de Caroline du Nord, nous avons également exploré la suppression des molécules HLA de classe I via la B-2-microglobuline (B2M) pour réduire le risque de rejet immunitaire. Une expression de HLA-ABC de 15,1±14,6% (N=11) a été obtenue après knockout par CRISPR/Cas9. Nous travaillons également sur la surexpression de HLA-E et HLA-G pour prévenir la lyse médiée par les cellules NK, nécessitant des optimisations supplémentaires.En conclusion, générer des HLA-E+ ou G+/B2M-/CAR-VST offre une alternative prometteuse pour créer des cellules entièrement allogéniques. Ces CAR-VST modifiés conservent leurs fonctions antivirales et antitumorales, ce qui en fait des candidats prometteurs pour les immunothérapies prêtes à l'emploi qui pourraient réduire les risques de rejet immunitaire et de GvHD
CAR-T cell therapy have revolutionized cancer treatment by modifying a patient's T cells to target specific tumor antigens. This personalized approach has shown remarkable success in treating B-cell malignancies like leukemia and lymphoma. However, the process is costly and time-consuming, as it involves collecting and modifying the patient's own cells, which delays treatment. Moreover, some patients may not have sufficient or viable T cells due to prior treatments or advanced disease stages, limiting the availability of CAR-T therapies for all patients.To address these challenges, allogeneic CAR-T cells from healthy donors provide a faster and more scalable solution, reducing production time and costs. However, these off-the-shelf therapies face risks like graft-versus-host disease (GvHD), where donor cells might attack the patient's tissues. Our study explored combining CAR technology with Virus Specific T cells (VSTs), known for their antiviral and antitumor properties, to generate CAR-VSTs. These dual-specific CAR-VSTs present a promising alternative, especially for patients prone to both tumor relapse and viral reactivation.In our study, we generated CAR-Ts and CAR-VSTs from same donors obtaining 40.28%±9.30% and 35.96%±11.40% CD19.CAR expression on day 7 (N=3), respectively. In vitro, CAR-VSTs showed robust tumor clearance similar to CAR-Ts, achieving 74.13%±22.06% lysis of CD19+ tumor cells. In a murine lymphoma model, both CAR-VSTs and CAR-Ts demonstrated comparable antitumor efficacy, successfully controlling tumor growth and improving survival. Moreover, CAR-VSTs maintained their antiviral function, efficiently lysing 62.32%±13.84% virus-peptide-pulsed cells, similar to native VSTs. We assessed the alloreactivity of CAR-VSTs and found that they exhibited significantly lower CD3 proliferation rates (28.27%±21.64%) compared to CAR-T cells (88.3%±24.48%, p=0.0285, N=4), indicating a reduced risk of GvHD. CAR-VSTs' dual-specificity for both tumor and viral antigens makes them a powerful tool to address cancer relapse and viral complications in patients.In collaboration with the University of North Carolina, we explored strategies to delete HLA class I molecules in CAR-VSTs by targeting B-2-microglobulin (B2M), aiming to reduce immune rejection. In addition, we worked on overexpressing tolerogenic molecules such as HLA-E and HLA-G to prevent NK cell-mediated lysis. Our results showed an HLA-ABC expression of 15.1±14.6% (N=11) after CRISPR/Cas9 knockout, which indicates successful deletion, though further optimization is necessary to prevent NK-lysis by re-expressing HLA-E or HLA-G.In conclusion, generating HLA-E+ or G+/B2M-/CAR-VSTs offers a promising alternative for creating fully allogeneic cells. These modified CAR-VSTs retain their dual antiviral and antitumor functions, making them a promising candidate for "off-the-shelf" immunotherapies that could reduce the risks of immune rejection and graft-versus-host disease