Preclinical evaluation and structural optimization of anti-BCMA CAR to target multiple myeloma.

Chimeric antigen receptor (CAR) T-cell based immunotherapy has become a promising treatment mainly for hematological malignancies. Following the major success of CD19-targeted CAR, new potential targets for other malignancies are required. As such, B-cell maturation antigen (BCMA) is an attractive tumor-associated antigen to be targeted in multiple myeloma (MM). Herein, we aimed at assessing the function and optimal configuration of different BCMA-specific CAR, based on the same targeting moiety but with a different hinge and co-stimulatory domain. We compared their function to that of a previously characterized BCMA-CAR used in clinical trials. All constructs were expressed at high levels by primary human T cells and could trigger cytokine production and cytotoxicity upon co-culture with multiple myeloma targets. Nonetheless, critical differences were observed in off-target activation, exhaustion, and activation marker expression and in vivo antitumoral activity mediated by these different constructs. Interestingly, we noted that CD8-based hinge, combined with a 4-1BB intracellular domain, proved superior compared to IgG4-connecting regions, and/or a CD28-signaling moiety respectively. Overall, this study emphasizes the influence of CAR primary structure on its function and led to the identification of a highly efficient BCMA-specific CAR, namely H8BB, which displayed superior anti-tumoral activity both in vitro and long-term in vivo efficacy.

Targeting glycosylated antigens on cancer cells using siglec-7/9-based CAR T-cells.

Chimeric antigen receptor (CAR) T-cells treatment demonstrate the increasing and powerful potential of immunotherapeutic strategies, as seen mainly for hematological malignancies. Still, efficient CAR-T cell approaches for the treatment of a broader spectrum of tumors are needed. It has been shown that cancer cells can implement strategies to evade immune response that include the expression of inhibitory ligands, such as hypersialylated proteins (sialoglycans) on their surface. These may be recognized by sialic acid-binding immunoglobulin-type lectins (siglecs) which are surface receptors found primarily on immune cells. In this regard, siglec-7 and -9 are found on immune cells, such as natural killer cells, T-cells, and dendritic cells and they can promote immune suppression when binding to sialic acids expressed on target cells. In the present study, we hypothesized that it is possible to use genetically engineered T-cells expressing siglec-based CARs, enabling them to recognize and eliminate tumor cells, in a non-histocompatibility complex molecule restricted way. Thus, we genetically modified human T-cells with different chimeric receptors based on the exodomain of human siglec-7 and -9 molecules and selected optimal receptors. We then assessed their antitumor activity in vitro demonstrating the recognition of cell lines from different histologies. These results were confirmed in a tumor xenograft model exemplifying the potential of the present approach. Overall, this study demonstrates the benefit of targeting cancer-associated glycosylation patterns using CAR based on native immune receptors and expressed in human primary T-cells.

MYC Induces Immunotherapy and IFNγ Resistance Through Downregulation of JAK2.

Immunotherapy has revolutionized the treatment of advanced melanoma. Because the pathways mediating resistance to immunotherapy are largely unknown, we conducted transcriptome profiling of preimmunotherapy tumor biopsies from patients with melanoma that received PD-1 blockade or adoptive cell therapy with tumor-infiltrating lymphocytes. We identified two melanoma-intrinsic, mutually exclusive gene programs, which were controlled by IFNγ and MYC, and the association with immunotherapy outcome. MYC-overexpressing melanoma cells exhibited lower IFNγ responsiveness, which was linked with JAK2 downregulation. Luciferase activity assays, under the control of JAK2 promoter, demonstrated reduced activity in MYC-overexpressing cells, which was partly reversible upon mutagenesis of a MYC E-box binding site in the JAK2 promoter. Moreover, silencing of MYC or its cofactor MAX with siRNA increased JAK2 expression and IFNγ responsiveness of melanomas, while concomitantly enhancing the effector functions of T cells coincubated with MYC-overexpressing cells. Thus, we propose that MYC plays a pivotal role in immunotherapy resistance through downregulation of JAK2.

Artifact3-D: New software for accurate, objective and efficient 3D analysis and documentation of archaeological artifacts.

The study of artifacts is fundamental to archaeological research. The features of individual artifacts are recorded, analyzed, and compared within and between contextual assemblages. Here we present and make available for academic-use Artifact3-D, a new software package comprised of a suite of analysis and documentation procedures for archaeological artifacts. We introduce it here, alongside real archaeological case studies to demonstrate its utility. Artifact3-D equips its users with a range of computational functions for accurate measurements, including orthogonal distances, surface area, volume, CoM, edge angles, asymmetry, and scar attributes. Metrics and figures for each of these measurements are easily exported for the purposes of further analysis and illustration. We test these functions on a range of real archaeological case studies pertaining to tool functionality, technological organization, manufacturing traditions, knapping techniques, and knapper skill. Here we focus on lithic artifacts, but the Artifact3-D software can be used on any artifact type to address the needs of modern archaeology. Computational methods are increasingly becoming entwined in the excavation, documentation, analysis, database creation, and publication of archaeological research. Artifact3-D offers functions to address every stage of this workflow. It equips the user with the requisite toolkit for archaeological research that is accurate, objective, repeatable and efficient. This program will help archaeological research deal with the abundant material found during excavations and will open new horizons in research trajectories.

Toward the identification of social signatures in ceramic production - An archaeological case study.

Ceramic analysis has been concerned with categorizing types according to vessel shape and size for describing a given material culture at a particular time. This analysis' long tradition has enabled archaeologists to define cultural units across time. However, going into the analysis of sub-typological variations is rarely done, although their meanings bear significant consequences on the understanding of ties between individuals and social units. This study, aiming to assess whether it is possible to identify social signatures, focuses on a single archaeological ceramic type. For this propose, we selected a corpus of 235 storage jars from two distinct periods: storage jars from the Intermediate Bronze Age (the 25th -20th century BCE); and the Oval Storage Jar type (hereafter: OSJ) from the Iron Age II (the late 9th-early 6th century BCE). The vessels selected were 3-D scanned to extract accurate geometric parameters and analyzed through an advanced shape analysis. The study results show that integrating computational and objective analysis methods, focusing on the "minute variation" within a single ceramic type, yields substantial insights regarding the relationship between variability and social units. In addition to the methodological guidelines and the suggested "work protocol" for further studies, the results shed light on the social organization of the Intermediate Bronze Age and the Iron Age II in Southern Levant.