Single Car Research Articles

Short-term noise annoyance towards drones and other transportation noise sources: A laboratory study.

Noise from unmanned aerial vehicles, commonly referred to as "drones," will likely shape our acoustic environment in the near future. Yet, reactions of the population to this new noise source are still little explored. The objective of this study was to investigate short-term noise annoyance reactions to drones in a controlled laboratory experiment. Annoyance to (i) two quadcopters of different sizes in relation to common contemporary transportation noise sources (jet aircraft, propeller aircraft, helicopters, single car passbys), and (ii) different drone maneuvers (takeoff; landing; high, medium, and low flybys) flown at different speeds and elevations was systematically assessed. The results revealed that, at the same sound exposure level, drones are perceived as substantially more annoying than other airborne vehicles and passenger cars. Furthermore, for drone maneuvers, landings, and takeoffs are more annoying than flybys, as are maneuvers flown at low speed. Different loudness metrics (LAE, LDE, effective perceived noise level, psychoacoustic loudness level) accounted for drone noise annoyance ratings to an equal degree. An analysis of psychoacoustic parameters highlighted the significant link between drone noise annoyance and tonality, sharpness, and loudness level. The results suggest a different perception and an increased annoyance potential of drones, which will likely require specifically tailored legislation.

Abstract B037: Banana Lectin expressing CAR T cells to target Pancreatic Cancer

Abstract To overcome both the heterogenous and dynamic expression of tumor antigen expression in pancreatic ductal adenocarcinoma (PDAC) and the physical barrier provided by the associated fibrotic stroma, we have developed a novel therapy that recognizes and binds to distinct patterns of aberrant glycosylation specifically expressed on tumor cells and their surrounding stroma. Aberrant glycosylation patterns are a hallmark of cancer and are widely present on tumors including PDAC. We therefore incorporated H84T BanLec as part of a chimeric antigen receptor and co-expressed the construct in T cells and showed that H84T BanLec CAR expressing T-cells can effectively target pancreatic tumors. We hypothesized that our novel lectin CAR would enhance the activity of T cells when co-expressed with conventional scFV – derived CARs targeting tumor associated antigens We compared the activity of dual-expressing H84T/HER.2 CAR-T or H84T/PSMA CAR-T cells against single- expressing CARs targeting PDAC. We determined anti-tumor cytotoxicity by measuring residual GFP+ tumor cells in a sequential killing assay using flow cytometry. We observed superior anti-tumor activity using dual expressing CAR T cells compared to T cells expressing either HER.2- or H84T- or PSMA- CAR alone. Similarly, in 3D multicellular spheroids containing tumor and cancer-supportive stromal cells, an IncuCyte live imaging assay shows dual CAR T cells have greater cytotoxicity against PDAC than single CAR expressing T cells (p= 0.02 H84T/HER.2 vs HER.2 CAR; p=0.00003 H84T/PSMA CAR vs PSMA CAR). These results also translate to improved anti- tumor function of dual CAR T cells in PDAC xenograft in vivo models where H84T/HER.2 CAR T cells significantly reduced the tumor volume burden (p=0.015; H84T/HER.2 average=57.1mm3; HER.2 average=116.1mm3; H84T average=130.3mm3). These findings suggest that H84T BanLec CAR T cells provide a tool to target the aberrant glycosylation patterns of malignant cells and their supporting stroma and may contribute to an effective treatment option for PDAC patients. We are now elucidating the underlying mechanism by which H84T BanLec CARs enhance tumor activity and promote T cell infiltration. Citation Format: Katie McKenna, Ada I Ozcan, David Markovitz, Malcolm K Brenner. Banana Lectin expressing CAR T cells to target Pancreatic Cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B037.

Resolution of Anterior Uveal Infiltration of Acute Lymphoblastic Leukemia After Chimeric Antigen Receptor T-Cell Therapy

Purpose: To present a case of acute B-cell lymphoblastic leukemia (B-ALL) ocular relapse treated with chimeric antigen receptor T-cell (CAR T) therapy. Methods: A retrospective case review was evaluated. Results: A 69-year-old woman with a history of B-ALL presented with bilateral hypopyons and 3+ anterior chamber cells that responded to topical prednisolone. Six months later, she developed recurrent hypopyons, iris neovascularization, and iris nodularity in both eyes. A fine-needle aspiration biopsy confirmed ocular infiltration of B-ALL. A single infusion of CAR T therapy resulted in remission of the systemic and ocular findings. The clinical course was complicated by cytokine release syndrome and neurotoxicity, which improved with medical management. The patient remained in remission for 7 months after a single CAR T infusion. Conclusions: CAR T therapy may be effective in treating systemic leukemia and uveal infiltration, with an ocular side-effect profile and visual outcomes. The systemic side effects of CAR T therapy may be managed medically.

Methods for ice crossings load capacity calculation

Introduction. The article deals with a problem relevant to the territory of our country – the calculation of the bearing capacity of ice crossings and winter roads. This problem is becoming more and more urgent in connection with the development of the northern territories, an increase in the carrying capacity of road transport and the volume of cargo traffic. At the same time, the construction of permanent roads and bridge crossings requires very large capital expenditures, which are not yet available for the Russian Federation.Methods and materials. A critical analysis of methods for determining by calculation the bearing capacity of ice cover at crossings (the maximum permissible load on the axle of a single car or road train) was carried out.The recommendations of normative and methodological documents on this issue are considered. The presented analysis of domestic and foreign publications made it possible to evaluate mathematical models of different levels of complexity and detail, with a different set of factors influencing the final result.Results. As a result of the research, a summary table, which contains formulas for calculating the bearing capacity of the ice cover, the parameters that are taken into account in these formulas, as well as the values of the bearing capacity calculated using these formulas for two temperatures: 0 °C and minus 20 °C has been compiled.Conclusions. According to the results of the analysis, it can be seen that the largest number of parameters is taken into account in M.M. Kazansky-R.A. Shulman; Q. Wang; ODM 218.4.030-2016 Methodological recommendations for assessing the carrying capacity of ice crossings’ dependencies. At the same time, the value of the bearing capacity, calculated according to the 11th formulas, varies by 2-3 or more times presented. Consequently, the presented mathematical models for predicting the carrying capacity of ice crossings require experimental verification on real objects by dragging the control cargo (with some refinement of this method).

IMMU-10. DEVELOPING NOVEL CHIMERIC ANTIGEN RECEPTOR THERAPIES FOR PAEDIATRIC HIGH GRADE GLIOMA

Abstract Paediatric high-grade gliomas (pHGG), particularly Diffuse Midline Glioma (DMG), present a formidable challenge in oncology. Due to the aggressive biology and location of pHGGs the implementation of adult treatment modalities (surgery, chemotherapy, and radiotherapy) is severely limited resulting in pHGG remaining a terminal disease. Thus, there is a pressing and urgent unmet clinical need for the development of innovative therapeutics. We will demonstrate the identification of cell surface targets using mass spectrometry proteomics. We have developed novel binders and engineered them into second-generation CAR T cells. We will present pre-clinical in vivo intracranial mouse models examining the efficacy of novel CAR T cell therapies. We have recently reported effective CART cell targeting of HER2 in DMG, and presently we will also demonstrate superior efficacy of additional CAR T cell targeting, both in vitro and in vivo in preclinical PDX models of DMG. We will present new data that CAR T cells targeting EphA3 show curative efficacy in pre-clinical models of DMG. Recognising that single targeting CAR T cells are unlikely to provide long-term clinical benefit, we will also present our combination approaches using CAR T cells targeting antigens such as GD2, EGFR, HER2, EphA3, CD276 and CD63. We are expanding our research to other CAR T combination strategies focusing on immune-modulating drugs such as checkpoint inhibitors or co-stimulatory agonists. We are committed to advancing CAR T cell therapies for pHGG patients and their families with an aim to aid in the development of much-needed new therapeutic options.

Prediction of brake pad wear and remaining useful life considering varying vehicle mass and an experimental holistic approach

Estimation of passenger car brake pad wear is considered, including real-time mass estimation to account for variations in vehicle mass. A longitudinal dynamics model is used to estimate mass, the application of the brakes, and a longitudinal braking force. Wear is modelled from brake work based on the product of the braking force and distance travelled during braking. Data is provided by standard vehicle on-board diagnostics data and a sensor fusion based on an inertial measurement unit and satellite position data, sampled from a single car with 10,200 km of on-road driving, including 31 brake pad wear measurement samples, and driving with up to 23% increased vehicle mass. The mass is accurately estimated within ±5% of the actual. Braking states determined from comparisons of measured and modelled deceleration provide accuracies of ≈91%, precisions of ≈96%, and recall rates of ≈86%, with inclusion of varying mass only resulting in minor differences. Brake work is estimated with (a) constant mass, (b) estimated mass, (c) actual mass, and (d) actual mass and braking states. Compared to (a), (b) and (c) provide 9% and 11% increased work, while (d) provides an increase of about 5% compared to (c). Wear coefficients are fitted on the initial half of the wear data considering (a), (b), and (d). Here (a) and (b) provide similar wear rates of k≈−1.22 mm/GJ, while (d) is about 5% lower. Validating on the second half of the data, (b) and (d) provide very similar wear estimates, with overall lower errors than (a). At the final sample, however, (a) provides a slightly lower absolute error, overestimating the wear by about 2%, while (b) and (d) underestimate it by about 3%. RUL, in terms of distance, is estimated across about 9,500 km of data by estimating a running ratio between cumulated work and distance. Here only (a) and (b) are considered, with (a) overestimating the RUL by 600 km at the actual wear limit, while (b) underestimates it by 190 km. A distance-based model is also considered, resulting in a constant RUL error of about 2,000 km.

ОБОСНОВАНИЕ КОНСТРУКЦИИ КРЫТОГО ГРУЗОВОГО ВАГОНА С УВЕЛИЧЕННЫМ ОБЪЕМОМ ГРУЗОВОГО ПОМЕЩЕНИЯ

The market of covered freight cars, as well as existing designs of this car type of domestic and foreign production is analyzed. Promising trends of making innovative designs of freight cars for the transportation of piece and container goods are found out. The original design of a covered freight car with an increased body space due to the use of the space between trucks is proposed. To reduce the complexity of loading and unloading operations, sliding sections of the side walls and roof are provided in the car design. To confirm the compliance of the proposed design with the requirements of regulatory and technical documents, the car was entered into the dimension, checking for automatic coupling at the junction of a straight line and a curve, passage in the coupling of an s-shaped curve and coupling of a straight line and a curve, and a single car along a circular curve of a small radius, as well as passing without self-decoupling of the hump of the sorting slide, the ramp exit of the train ferry. A safety assessment is carried out with regard to rolling the wheel onto the rail and overturning the car in curves. The car dynamic characteristics are found out by mathematical modeling methods. The assessment of the strength of the body load-bearing structure, lifting mechanisms of the floor sections, sliding sections of the roof and side walls is performed using the finite element method implemented in the environment of a specialized software package. The results obtained as a result of calculations and computer modeling confirmed the operability and safety of the proposed design of a covered freight car with an increased loading space.

Evaluation of Car-Following Model for Mixed Autonomous and Human Driven Vehicles on Road Facilities.

Driverless cars are emerging slowly but bear the opportunity to improve the traffic system efficiency and user comfort. For the near future, a mix of human-driven and driver-less vehicle co- existence is inevitable. The quest to integrate autonomous and human-driven vehicles has created numerous critical questions in the road traffic system: How can autonomous and human- driven vehicles co-exist efficiently? What are the prospects of breaching the existing car-following model widely adopted to control the longitudinal movement of vehicles and control the driver’s maintenance of a safe distance? There is no doubt, that the emergence of autonomous vehicles and their integration is altering the existing traffic theories and the idea behind the car-following model. Currently, researchers are working hard to efficiently describe the physics behind the traffic flow theory of car-following models in mixed traffic. This work investigated a homogeneous car-following model sensitivity with varying human drivers’ reaction time in mixed traffic. The ability to predict the optimal vehicle response time in a platoon to the behaviour of its predecessor is vital in predicting the effect a change in the driving environment of mixed traffic has on traffic flow. The major complexities surrounding the car- following model in a mixed traffic flow environment are how the traffic participants share the road intersection space and the car-following behavior of human-driven vehicles. Investigation in microscopic traffic flow describes the way a single car be- haves and then introduces the impact another car has on a car’s behavior to address the mixed traffic flow. The traffic speed performance under the different traffic conditions suggests that the proposed mixed traffic car-following model outperforms the existing car-following model for human-driven vehicles with an average of 22.59mph. There is a significant throughput improvement in a mixed traffic environment when the distance headway of autonomous vehicles is adjusted to support the research hypothesis.

Kinematic simulations and design of a steering upright for a single seater electric car

Kinematic simulations and design of a steering upright for a single seater electric car

A Compilation of Experiences in Utilizing CAR-T Cell Therapy for Richter's Transformation

Objective:Even with Bruton tyrosine kinase inhibitors (BTKis) and BCL2 inhibitors, the prognosis for Richter's Transformation (RT) remains dismal, with a median overall survival (OS) of only 9-12 months from initial diagnosis. Although there are reports on CAR-T therapy for RT, the sample size is small and the conclusions vary greatly. Methods: We conducted a retrospective investigation of the efficacy of CAR-T cell therapy in treating RT at our center. Each participant provided their informed consent to partake in the “Phase II Clinical Trial of Sequential CAR-T Cell Therapy Targeting Different B-Cell Epitopes for Recurrent, Refractory Aggressive B-Cell Lymphoma”(Protocol number: BR-XG-CART-II, ChiCTR2100055062, Ethical Approval Number 20191225-PJ-003). Before lymphocyte collection, chemotherapy regimen without fludarabine/ Bendamustine is first used to reduce peripheral blood tumor load if peripheral blood tumor cells are higher than 10%. Patients with extramedullary tumor loads of 5cm or larger were treated with tumor reduction chemotherapy between lymphocyte collection and CART cell transfusion. The pretreatment scheme based on Bendamustine or FC scheme was adopted before CAR-T cells were infused. The efficacy and safety of CART cells were observed. Results: Between Feb 1, 2020 and Feb 28, 2023, our center administered CAR-T cell therapy to 16 patients diagnosed with RT, all of whom developed into diffuse large B-cell lymphoma. The patient population had a median age of 59 years (range: 42-69 years), consisting of 62.5% (10/16) males and 37.5% (6/16) females. The ECOG scores varied between 0 and 3. The median number of prior treatment lines was 5 (2-7). 93.8% failed previous BTKis treatment and 75% failed BCL2 inhibitor treatment; 93.8% had undergone chemotherapy (median 6 courses). 1 patient had previously failed universal CAR-T cell therapy, and another relapsed after autologous transplantation. 90% had at least one TP53 abnormality detected by either FISH or second-generation gene sequencing. Extramedullary masses were found in 87.5% (14/16) of patients, with a median maximum diameter of 42.5mm (18-144mm). Masses larger than 5cm were found in 25% (4/16) of cases, marrow involvement was noted in 62.5% (10/16), and cerebrospinal fluid was affected in 18.75% (3/16). None of the patients had central masses observed in their MRIs. During lymphocyte collection, 58.3% of patients showed positivity for peripheral blood tumor cells, with a maximum tumor cell proportion of 5.69%. The median infused CAR-T cell dose was 1.37×10 6/kg. 12 of them were injected with mouse CD19CAR-T, 2 with human CD20CART, and 2 with human CD19CART. CRS occurred in 81.3%of patients, of which, 68.8%were grade 1-2 and 12.5%were grade 3. Central CRS occurred in 6.3% (1/16) of patients. The complete response (CR) rate at the 3-month mark stood at 56.3% (9/16), with an overall response rate (ORR) of 68.8% (11/16). As of July 6, 2023, the median follow-up time was 10.6 months (4.4-33.6m), the 1-year OS was 73.4%, and the 1-year PFS was 68.8%. Median OS and PFS were not reached. One of the five patients who did not respond later underwent allogeneic hematopoietic stem cell transplantation and is still alive. The other four, despite attempts at salvage treatment including second CAR-T cell therapy, all died, with a median OS of only 5.6 months (4.4-8.9m). None of the four patients with extramedullary masses larger than 5cm achieved ORR despite tumor-reducing chemotherapy prior to CAR-T infusion. Only 1 of 3 patients with central involvement achieved CR and experienced grade 3 CRS along the way, while the other 2 did not respond to CAR-T therapy. Among all patients, only one experienced central CRS, and his cerebrospinal fluid showed no signs of tumor cells prior to the administration of CAR-T treatment. A patient who was in CR succumbed to an acute coronary event 19 months following infusion, despite remaining in CR. Conclusion: Even in the presence of TP53 gene abnormalities, resistance to BTK/BCL2 inhibitors, and unsuccessful previous chemotherapy regiments, A single CAR T cell therapy can result in CR in more than half of RT patients. Nonetheless, the prognosis remains bleak for patients presenting with large tumor masses and central nervous system involvement, despite having received chemotherapy aimed at reducing tumor size. The outlook is particularly dismal for those patients who fail to respond to CAR-T cell therapy.

The Clinical Outcome of Patients with Refractory/Relapsed Lymphoid Blastic Phase of Chronic Myeloid Leukemia Treated with CAR T-Cells Therapy

Introduction : Chimeric antigen receptor T (CAR T) is an effective therapy for refractory/relapsed (R/R) philadelphia chromosome positive acute lymphoblastic leukemia (ph+ ALL) patients in TKIs era. Patients with lymphoid blastic phase of chronic myeloid leukemia (CML-LBC) also have BCR-ABL fusion oncogenes, and usually shows an aggressive clinical course with a poor prognosis. However, outcomes of CAR T therapy for patients with R/R CML-LBC is still uncertain. This study was designed to compare the efficacy and safety of CAR T-cells therapy in patients with R/R CML-LBC and ph+ ALL. Patients and Methods : Clinical data from 13 patients with R/R CML-LBC and 121 R/R ph+ ALL patients who received CAR T treatments at our center from February 2017 to March 2023 were retrospectively collected and analyzed. Before CAR T-cells treatment, fifty patients (11.2%) had primary refractory disease, 106 patients (79.1%) were experiencing the first relapse, and 13 patients (9.7%) were experiencing the second or more relapse. All patients received lymphodepletion with fludarabine (30 mg/m2 /d) and cyclophosphamide (300 mg/m2 /d) -based conditioning regimens on day −5 to −3. CAR T-cells were then infused at day 0. Among them, 8 patients with CML-LBC and 92 ph+ ALL patients received single CD19 CAR T therapy, others received tandem CD19/CD22 CAR T. Results:The baseline characteristics of all patients were shown in Table 1. Patients with CML-LBC carried a higher proportion of mutations in the ABL kinase region ( P=0.036).The clinical timeline of CML-LBC patients as shown in Figure 1A,and ABL kinase region mutations and gene mutations of patients are shown in Figure 1B. Patients with CML-LBC (58.3%) had significantly lower CR rates than ph+ ALL (97.2%) patients at +2 month after CAR-T therapy (P<0.001) ( Fig. 1B). ABL kinase region mutation (odds ratio, 0.091; 95% CI, 0.015-0.571) and CML-LBC (odds ratio, 0.063; 95% CI, 0.010-0.376) remained risk factors in multivariate logistic regression analysis of CR rate in all patients. Furthermore, minimal residual disease (MRD)-negative CR rates were 58.3% and 95.3% at +2 month after CAR T-cells therapy, respectively (P<0.001) ( Fig. 1B). There was no significant difference in the incidence of adverse events among the two groups. Severe cytokine release syndrome (CRS, Grade ≥ 3) occurred in 7.7% of patients with CML-LBC and 8.3% in ph+ ALL patients (P=1.000). Moreover, patients with CML-LBC showed poorer 2-year prognoses than the ph+ ALL patients (overall survival: P=0.0068; leukemia-free survival: P<0.0001; cumulative incidence of relapse: P<0.001) ( Fig. 1C-1E). Univariate and multivariate Cox regression analyses showed that a poorer LFS was related to ABL kinase region mutation (hazard ratio, 3.257; 95% CI, 1.404-7.553) and CML-LB (hazard ratio, 3.803; 95% CI, 1.507-9.596). Conclusions: Worser sustained antitumor efficacy and long-term survival with CAR T-cells therapy in patients with CML-LBC compared to ph+ ALL patients, and ABL kinase region mutation is an independent risk factor for CR and LFS.

CD33-TIM3 Dual CAR T Cells: Enhancing Specificity While Maintaining Efficacy Against AML

Background: The myeloid associated antigen CD33 is overexpressed in over 88% of Acute myeloid leukemia (AML) patients, making it a suitable target antigen for CAR T cell therapy across different genetic subtypes. Despite the development of clinical trial data, there remains a concern about on-target off-leukemia toxicity. To enhance specificity, we generated dual targeting CAR T cells that target CD33 and/or T-cell immunoglobulin and mucin-domain containing-3 (TIM3). The latter is known to be expressed on LSCs, but not on healthy hematopoietic precursor or stem cells. Notably, TIM3 is recognized for its inhibitory immunomodulatory role further enhancing the suitability of this target antigen. The objective of this research was to explore various CD33 and TIM3 dual-targeting CAR T cells approaches that enhance AML specificity and maintain high anti-AML activity. Methods: The hybridoma technology was used to generate antibodies against TIM3 immunized mice. Antibodies were screened for TIM3 specificity via ELISA and FACS. The anti-TIM3 single-chain variable fragment (scFv) DNA sequence was sequenced from hybridoma cells. Furthermore, the binding ability to TIM3 antigen (PDB ID: 5F71) was evaluated using AlphaFold. The CD33 scFv was derived from Gemtuzumab ozogamicin (clone: hP67.6). ScFv sequences and co-stimulation domains (CD28 or 4-1BB) were cloned into a pMP71 vector. Retrovirus for transduction was produced using the 293Vec-GALV and RD114 retroviral production system. In vitro co-culture assays of CAR T cells and target cells were performed to study the efficacy of CAR T cells. The cytotoxicity against wild-type and TIM3 transduced AML cell lines (THP-1 and OCI-AML3) was assessed by multiparameter flow cytometry (MPFC). The secretion of effector cytokines (IFN-γ, TNF, IL-2) was analyzed via CBA assays. In addition, avidity between CAR T cells and target cells was determined by Z-Movie analyzer. The different CAR constructs were screened for on-target off-tumor toxicity in colony forming unit assays (CFU) using isolated CD34 + hematopoietic stem and progenitor cells (HSPCs) from healthy doners after 14 days. In order to compare CAR constructs regarding long-term efficacy in antigen restimulation assays, CAR T cells were co-cultured with irradiated TIM3 transduced OCI-AML3 every 4 days at an E: T ratio of 1:1 for 24 days. Moreover, CAR T-cell proliferation, checkpoint marker expression and T-cell subset differentiation were analyzed via MPFC. Results: All dual CAR T cells (compound, split, tandem, pooled) were generated with high and robust transduction efficacy (Figure 1A). TIM3-dependent fratricide was not observed during CAR T cell manufacturing. Compared to single antigen targeting CAR T cells, dual CAR T cells showed enhanced cytotoxicity against AML cell lines and primary AML cells (Figure 1B). Moreover, we also observed a strong increase in the secretion of proinflammatory cytokines (IFN-γ and IL-2) and higher avidity of dual CAR T cells. Notably, split CAR T cells demonstrated greater specificity in cocultures directed against mono vs. dual-target antigen expression cell lines. In accordance with these observations, split CAR T cells did not exert on-target off-leukemia toxicity against healthy HSC in CFU assays. In the antigen restimulation assay, we observed that compound CAR T cells exhibited diminished expanding capacity and heightened expression of exhaustion markers in comparison to the other CAR T constructs. Conclusion: We developed multiple CD33 and TIM3 dual CAR T cells using both “AND” and “OR” gating strategies in this study. Our findings revealed that dual CAR T cells provided higher avidity and cytotoxicity compared to single targeting CAR T cells against dual antigen expressing target cells in vitro. Importantly, only the split CAR T constructs demonstrated specific killing of CD33 +TIM3 + cell lines and primary AML cells while sparing HSPCs. Prospectively, compound, tandem, pooled CD33 TIM3 specific CAR T cells might be helpful in a bridge to transplant setting, whereas split CAR T cells might provide a higher safety profile, thereby allowing a transplant independent approach. To advance our concepts, we are currently conducting in vivo experiments in an NSG mouse model.

Development of a Phase 1 Study Evaluating the Activity of Modular CAR T for Multiple Myeloma (MCARTY) Targeting BCMA and CD19 for Improved Persistence

CONCLUSION BCMA directed CAR T cell therapy for multiple myeloma (MM) shows promise, but very long-term survival may be limited. Notably, BCMA CAR T persistence in MM is shorter than that seen in with CD19 CAR T cells in B cell malignancies where long-term CD19 CAR T persistence is associated with durable responses. Shorter BCMA CAR T persistence may be explained by the lower surface density of BCMA on tumour compared with that of CD19. Alternatively, CD19 CAR T persistence may be driven by recognition of continuous emergence of newly differentiated B cells. We sought to improve BCMA CAR T persistence by (a) developing a BCMA CAR with high sensitivity to BCMA, and (b) co-expression with a CD19 CAR. Notably, while CD19 is absent or expressed at low level on myeloma cells, CD19 may be expressed on a subpopulation of myeloma stem cells, providing an additional rationale for co-targeting CD19 in MM. We performed functional screening of several mid to high affinity BCMA binders in CAR format. We additionally explored whether a more stable CAR format would improve sensitivity. We explored co-expression of the new BCMA CAR with the CD19 CAR from obecabatagene autoleucel (also known as CAT19), which is known to drive long-term persistence. We also designed and started a clinical study (MCARTY, NCT04795882) testing the BCMA CAR alone or co-expressed with a CD19 CAR in patients with refractory Myeloma. The binding affinity for BCMA was determined for 13 antibody clones from a rat library, before being assessed functionally in single chain variable fragment (scFv) and fragment antigen-binding (Fab) formats in a second generation (41bbζ) CAR backbone. Noting superior target binding, functional efficacy and minimal basal activation of effector cells expressing the D8 binder in Fab format we then assessed this construct against standard BCMA targeting CARs based on the bb2121 and LCAR-B38M binders used in Ide-Cel and Cilta-Cel respectively. Compared to these competitor CARs, D8 demonstrated improved sensitivity to BCMA with improved activation, target kill, and cytokine release in a series of cocultures with plate bound antigen and cell targets expressing low level BCMA. Successful manufacture in an academic setting of dual targeting CAR T cells was achieved by double transduction of autologous CD4/CD8 selected T cells with separate lentiviral vectors titrated for equal transduction. Manufacture by CliniMACS Prodigy included AKT inhibitor VIII to reduce T cell differentiation and LentiBoost, a polaxamer-based transduction enhancer, in a protocol lasting 6-8 days. This resulted in 3 populations of CAR T cells (D8+CAT19-, D8-CAT19+, D8+CAT19+) which can be individually tracked in treated patients. The MCARTY trial is a single centre, Phase 1 dose escalation trial uniquely designed with two cohorts to evaluate the safety of D8 CAR T cells (Cohort 1) and D8/CAT19 CAR T cells (Cohort 2) in patients with relapsed/refractory MM at a dose of 50 or 150x10^6 CARs. Inclusion criteria are: progression or stable disease to last treatment, 3 or more previous lines of therapy, exposure to proteasome inhibitors (PI), immunomodulatory drugs (IMID) and CD38 antibodies, ECOG 0/1. There was no selection based on previous exposure to BCMA targeting agents or tumour BCMA expression. As of 11 July, 8 patients have been treated, 6 patients with D8 CAR alone (n=3 at each dose level) and 2 patients with the lower dose of dual targeting D8/CAT19 CAR T cells. Median age was 57 (range 33-66), high risk cytogenetics in 4 of 7 patients (57%), previous lines of therapy 4 (range 3-8). Doses were well tolerated with no cases of ICANS or CRS≥ grade 3. At a median follow up of 4.5 months (range 1-12), clinical responses ≥PR were seen in all patients (ORR: 100%). Of 6 patients, 5 (83%) CR/sCR, 1 (17%) PR at 3 months. We observed robust expansion of D8 CAR in Cohort 1, and of dual and single targeting CARs in Cohort 2. We report a unique approach to prolong CAR persistence in MM by developing a CAR T cell co-expressing a sensitive BCMA CAR construct and CD19 CAR with proven durability. We report feasibility of double transduction, the first clinical use of a FabCAR and a unique trial design allowing comparison of treatment with single vs dual targeting CAR constructs.

New Tocilizumab (TCZ) Dosing Guidance for T-Cell Engaging Bispecific Antibody-Related Cytokine Release Syndrome (CRS) in Patients (pts) with Relapsed/Refractory (R/R) B-Cell Non-Hodgkin Lymphoma (B-NHL): Insights from Pooled Clinical Trial Safety Experience and Quantitative Clinical Pharmacology (qCP) Analyses

Background: Mosunetuzumab (Mosun), approved for treatment of pts with R/R follicular lymphoma (FL) and ≥2 prior therapies, and glofitamab (Glofit), approved for treatment of pts with R/R diffuse large B-cell lymphoma, or large B-cell lymphoma arising from FL, and ≥2 prior therapies, are CD20xCD3 T-cell engaging bispecific antibodies that redirect T cells to eliminate B cells. CRS is a common on-target adverse event observed with bispecific antibodies and associated with exposure. TCZ, a humanized anti-interleukin-6 (IL-6) receptor, is often used for CRS management. TCZ dosing recommendations derived from CAR T-cell studies (Le et al. Oncologist 2018) may not be optimal for bispecific antibody-treated pts. Using TCZ clinical usage patterns in Mosun- or Glofit-treated pts with R/R B-NHL, qCP, and cytokine analyses, we explored a new TCZ dosing regimen for CRS management (for pts >30 kg body weight). Methods: Pooled safety data from seven Phase I/II studies evaluating Mosun or Glofit in pts with R/R B-NHL were analyzed. CRS was graded using ASTCT criteria (Lee et al. Biol Blood Marrow Transplant 2019). Modeling was used to predict TCZ concentrations (conc) and soluble IL-6 receptor (s-IL6R) occupancy over time, following administration of different TCZ dosing regimens in a subset of pts. Downstream cytokine pharmacodynamic responses (IL-6 and C-reactive protein [CRP]) were evaluated graphically post-TCZ dosing. Results: Data from 733 Mosun- and 772 Glofit-treated pts across different Mosun/Glofit dose levels were analyzed; of these, 232 (31.7%) and 427 (55.3%) pts, respectively, had ≥1 CRS event, and 36/232 (15.5%) and 140/427 (32.8%) pts received TCZ for CRS management. CRS events were mainly Grade 1/2 and occurred during the first treatment cycle. Most pts who received TCZ for CRS received ≤2 doses of TCZ overall (Mosun: 94.4%; Glofit: 84.2%) ( Table), and 1 dose of TCZ for the management of a single CRS event (Mosun: 91.7%; Glofit: 84.9%). CRS resolution was achieved within 3 days of TCZ administration in 75.0% and 66.9% of Mosun- and Glofit-treated pts, respectively. By then, a reduction in IL-6 and CRP levels was also observed. In a prior pharmacokinetic (PK) analysis of TCZ in pts with CAR T-cell-associated CRS, predicted TCZ conc after administration of 4 intravenous (IV) TCZ doses of 8 mg/kg every 8 hours (q8h) was generally below 649 μg/mL, the observed maximum conc in healthy volunteers and “safety threshold” used by the FDA (Le et al. Oncologist 2018). qCP analyses of TCZ PK and s-IL6R data from 67 pts who received TCZ for CRS management following treatment with Mosun or Glofit indicated that this TCZ regimen may exceed the safety threshold, in part because maximum TCZ conc is one-third lower in pts with CAR T-cell-induced CRS vs bispecific antibody-induced CRS. In addition, typical bispecific antibody regimens involve multiple doses compared with a single CAR T-cell infusion, with the potential for multiple CRS events requiring TCZ treatment; therefore, TCZ accumulation must be considered. qCP simulations predicted that for 1 x 8 mg/kg dose, median duration of >90% s-IL6R saturation was 28 (range: 14-28) days ( Table). Furthermore, simulations indicated that ≤2 consecutive TCZ doses q8h per CRS event and a maximum of 3 TCZ doses within a 6-week time period maintain the TCZ conc below the FDA safety threshold, while ensuring sustained s-IL6R occupancy ( Table). Conclusions: qCP results and clinical experience in Mosun and Glofit studies support a TCZ dosing regimen of ≤2 IV doses of 8 mg/kg per CRS event administered q8h, not exceeding a total of 3 doses within a 6-week period. Further improvement in CRS management and symptom resolution with additional TCZ doses is unlikely given the predicted duration of s-IL6R occupancy and observed rapid decline in systemic inflammatory markers post dosing. This proposed TCZ dosing regimen reflects TCZ usage patterns in clinical trials evaluating Mosun and Glofit, and is included in their Summaries of Product Characteristics. In patients with R/R B-NHL receiving bispecific antibody treatment, TCZ is generally suitable for management of acute CRS, and balances risk-benefit for potential scenarios involving multiple TCZ administrations for pts with CRS recurrence. This recommendation considers TCZ safety factors, the unique PK properties of TCZ in this setting and the expected duration of activity based on pharmacological modeling of target binding (s-IL6R occupancy).

A strategy for lightweight designing of a railway vehicle car body including composite material and dynamic structural optimization

AbstractRolling stock manufacturers are finding structural solutions to reduce power required by the vehicles, and the lightweight design of the car body represents a possible solution. Optimization processes and innovative materials can be combined in order to achieve this goal. In this framework, we propose the redesign and optimization process of the car body roof for a light rail vehicle, introducing a sandwich structure. Bonded joint was used as a fastening system. The project was carried out on a single car of a modern tram platform. This preliminary numerical work was developed in two main steps: redesign of the car body structure and optimization of the innovated system. Objective of the process was the mass reduction of the whole metallic structure, while the constraint condition was imposed on the first frequency of vibration of the system. The effect of introducing a sandwich panel within the roof assembly was evaluated, focusing on the mechanical and dynamic performances of the whole car body. A mass saving of 63% on the optimized components was achieved, corresponding to a 7.6% if compared to the complete car body shell. In addition, a positive increasing of 17.7% on the first frequency of vibration was observed. Encouraging results have been achieved in terms of weight reduction and mechanical behaviour of the innovated car body.

Intratumoral pan-ErbB targeted CAR-T for head and neck squamous cell carcinoma: interim analysis of the T4 immunotherapy study

BackgroundLocally advanced/recurrent head and neck squamous cell carcinoma (HNSCC) is associated with significant morbidity and mortality. To target upregulated ErbB dimer expression in this cancer, we developed an autologous CD28-based...

Canine osteosarcoma as a solid tumor model for evaluation of dual valent B7H3-CXCR2 CAR T cell therapy

Abstract To develop more effective solid tumor CAR T cell therapies, we utilized the spontaneous canine metastatic osteosarcoma (OS) animal model. Our studies evaluated the relative activity of single B7H3 and dual-valent B7H3-CXCR2 CAR constructs. Studies were also designed to determine if modifying the tumor microenvironment (TME) with oral immunotherapy improved CAR activity, based on recent studies of losartan and propranolol in dogs with OS and glioma. We evaluated B7H3 expression on canine OS cell lines using flow cytometry and OS biopsies using IHC. High levels of B7-H3 were observed on OS cell lines and OS biopsies, whereas normal canine tissues were negative or low. To assess the utility of the dual CAR we determined that canine OS cell lines secreted high IL8 spontaneously. To assess the functionality of canine B7-H3 CAR T cells, we generated canine CAR T cells from whole blood from dogs with cancer. We next determined that canine dual CAR T cells exerted specific killing and cytokine activity against B7H3+ canine OS cell lines. To further establish functionality, we evaluated the two CAR constructs in a mouse xenograft model of canine OS. We found that the dual CAR construct was significantly more active than the single CAR construct in slowing tumor growth. We found that losartan and propranolol blocked migration of mouse inflammatory monocytes to xenografted canine OS tumors. Future steps will be to evaluate the effectiveness of dual CAR T cells with TME modification in dogs with metastatic OS, to investigate the translational relevance in human patients. Results from these studies in dogs will likely give more clinically relevant results for evaluating new CAR T cell therapies and bridging results between mouse models and human clinical trials. The V Foundation, The Shipley Family Foundation, and NIH T32 training grant

Radar Signal Processing Architecture for Early Detection of Automotive Obstacles

With the mass adoption of automotive vehicles, road accidents have become a common occurrence. One solution to this problem is to employ safety systems that can provide early warning for potential accidents. These systems alert drivers to brake or take active control of a vehicle in order to make braking safer and smoother, thereby protecting drivers and all other road traffic participants. Most such safety systems utilize millimeter-wave radar as primary sensors, and one of the main challenges is real-time data processing from multiple sensors integrated into a single passenger car. When an obstacle is too close to a vehicle, often there is insufficient time to run higher-order digital signal processing algorithms; hence, the decision to brake must be made based on low-level hardware processing only. For that purpose, a hardware generator for the early detection of automotive obstacles that does not impede the operation of higher-order signal processing algorithms is described. The proposed generator is captured in the Chisel hardware design language and a method for reducing the overall ranging latency is presented. The system constraints are calculated using an exemplary radar front-end and the proposed generator parameters. The obtained analytical results are experimentally confirmed with a prototype composed of a typical industrial radar front-end while the signal processing back-end instance of the described generator was implemented on an FPGA board. The measurements demonstrate that with the fast proximity alert, objects can be detected in less than a hundred microseconds, thus considerably reducing the system reaction delay and braking distance.

Abstract 1148: 1XX mutations slow CAR T cell signaling and increase in vivo persistence

Abstract Purpose: Develop CAR T cells that have increased persistence in vivo and kill low antigen expressing solid tumor cancer cells. Experimental: Three CARs were generated that were all targeted to the tumor with the same antibody fragment, huMNC2, that recognizes a cryptic epitope on the MUC1 cleavage product called MUC1* (muk 1 star). One construct comprises a 41BB co-stimulatory domain, while the second construct has a CD28 co-stimulatory domain. The first 2 CARs have a wild-type CD3z, whereas the 3rd bears Michel Sadelain’s “1XX” mutations in CD3z. These are 2 Tyrosine to Phenylalanine mutations in ITAM 2 and ITAM 3. The 1XX mutations were designed to slow signaling to eliminate the problem of CAR T cell exhaustion. Human T cells were separately transduced with the 3 CARs and tested in NSG mice bearing human breast tumors. The tumors were heterogeneous tumors comprising parent cells that fluoresce red and express medium to low amounts of the antigen, MUC1*, and cells that fluoresce green wherein the parent cells have been transduced to express more MUC1*. The percent of the tumor that expressed high levels of MUC1* was varied from 7.5% to 15% to 30%. 150 female NSG mice were implanted with 500,000 tumor cells. The animals received a single CAR T cell injection of one of the three CAR T cells at an Effector to Target ratio of 10:1, 5:1, or 1:1. The experiment was allowed to proceed for 96 days, with periodic IVIS measurements to track tumor growth with a focus on tumor recurrence. After sacrifice, residual tumors were excised, dissociated and analyzed to determine if tumor recurrence was being driven by the high antigen cells or the low antigen cells. CAR T cells were recovered from the blood and the spleen. They were enumerated and analyzed by flow cytometry for the presence of markers of exhaustion. Unpublished Results: Tumor recurrence occurred in mice treated with CAR T cells bearing wild-type CD3z, but essentially did not recur in the mice treated with CAR T cells bearing the 1XX mutations. Four hundredfold more CAR T cells were recovered from the mice treated with the CAR-1XX cells compared to CARs with wild-type CD3z, and the CAR-1XX cells appeared not to be exhausted even after 96 days. CAR-1XX T cells administered at a 1:1 E:T ratio, with only 7.5% of the tumor comprised of high antigen expressing cells, were effectively killed with no significant tumor recurrence. In contrast, the CARs with wild-type CD3z had no effect on tumor growth in this group. Surprisingly, analysis of excised tumors showed that tumor recurrence in mice treated with CAR wild-type CD3z T cells was driven by the low antigen density cells, whereas the CAR-1XX T cells killed high antigen and low antigen density cells at the same rate. Conclusions: 1XX mutations in CD3z greatly increase CAR T persistence in vivo and unexpectedly enable the killing of low antigen tumor cells. Citation Format: Cynthia C. Bamdad, Benoit J. Smagghe, Mark G. Carter, Danica M. Walkley, Jac-Leen S. Nash, Kevin R. Yi, Trevor J. Grant, Laura M. Reale, Michael J. Nash, Gregory L. Riley, Andrew K. Stewart. 1XX mutations slow CAR T cell signaling and increase in vivo persistence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1148.

Behavioural norms or personal gains? – An empirical analysis of commuters‘ intention to switch to multimodal mobility behaviour

Existing research has revealed that there is a growing dominance of multimodality in urban areas, which is frequently considered to be an important part of the solution to the burdens associated with heavy car use, such as traffic congestion, noise, and air pollution, as well as associated risks to human health. Yet, while the role of private motorized transport is decreasing in densely populated urban areas, monomodal travel patterns, as exemplified by single occupancy car use, are still dominating work-related mobility. To date, studies on modal shift have investigated the decision-making processes with regard to particular travel modes. However, the intention to switch to multimodal mobility in a commuting context, which may represent a first step towards car use reduction, remains relatively unexplored. Therefore, this paper aims to understand and identify the predecessors of car commuters’ intention to switch to multimodal commuting via an online questionnaire. Our theoretical framework is based on an extension of Ajzen’s Theory of Planned Behaviour (TPB) with Lindenberg and Steg’s Goal-framing Theory (GFT), including a novel fourth goal frame, habit, and person-organisation fit (POF) as additional constructs. Contrary to our expectations, gain and hedonic goals, as well as POF and habit, were not found to affect switching intention. Instead, our findings suggest that a switch to multimodal mobility is strongly dependent on people's normative goals, underlining the need for employers to encourage and reward sustainable multimodal commuting. These findings can assist policy makers, corporate mobility, and HR managers to promote more sustainable, multimodal mobility behaviour.