ZFP36 disruption is insufficient to enhance the function of mesothelin-targeting human CAR-T cells.

Loss of inflammatory effector function, such as cytokine production and proliferation, is a fundamental driver of failure in T cell therapies against solid tumors. Here, we used CRISPR/Cas9 to genetically disrupt ZFP36, an RNA binding protein that regulates the stability of mRNAs involved in T cell inflammatory function, such as the cytokines IL2 and IFNγ, in human T cells engineered with a clinical-stage mesothelin-targeting CAR to determine whether its disruption could enhance antitumor responses. ZFP36 disruption slightly increased antigen-independent activation and cytokine responses but did not enhance overall performance in vitro or in vivo in a xenograft tumor model with NSG mice. While ZFP36 disruption does not reduce the function of CAR-T cells, these results suggest that singular disruption of ZFP36 is not sufficient to improve their function and may benefit from a multiplexed approach.

Understanding melanoma in situ: Lentigo maligna surgical treatment terminology and guideline adherence, a targeted review.

BACKGROUND: Malignant melanoma in-situ, lentigo maligna (MMIS-LM) can be successfully treated with several different surgical techniques; however, the literature is inconsistent in defining them. OBJECTIVE: To comprehensively define and describe the national guideline recommended surgical techniques used to treat MMIS-LM to help clarify and standardize this terminology to ensure compliance with the guidelines. METHODS: A targeted literature review was performed from 1990 to 2022 focusing on articles that discussed the national guideline recommended surgical techniques of wide local excision, Mohs micrographic surgery (MMS), modified Mohs surgery, and staged excision/Slow-Mohs for MMIS-LM, as well as the related methods of tissue processing. National Comprehensive Cancer Network and American Academy of Dermatology guidelines were reviewed to identify how the techniques need to be employed to be compliant with guideline recommendations. RESULTS: We describe the various surgical and tissue processing techniques and discuss advantages and disadvantages of each. LIMITATIONS: This paper was styled as a narrative review defining and clarifying terminology and technique and does not investigate these topics more broadly. CONCLUSION: Understanding the methodology and terminology for these surgical procedures and tissue processing methods is critical so that both general dermatologists and surgeons can employ these techniques effectively for optimal patient care.

Structural Differences in Meibum From Donors After Hematopoietic Stem Cell Transplantations.

PURPOSE: Meibum is considered to be a key component of tears that serve to protect the eye, and conformational changes in meibum have not been studied extensively within the population of patients who had hematopoietic stem cell transplantation (HSCT). The aim of this study was to determine possible lipid conformational changes in the meibum of patients who had HSCT. METHODS: Participants who had HSCT were randomly sampled for this prospective comparative study. Control participants did not have dry eye or had not undergone allogeneic or autologous stem cell transplantation. Fourier-transform infrared spectroscopy was used to measure meibum phase transition. RESULTS: Meibum was collected from both eyes of 36 donors without dry eye (Mc) and from 22 patients who had undergone HSCT (MHSCT). There were no significant differences between the phase transition parameters based on gender or race. The following were the significant differences (P < 0.0001) between the parameters for Mc compared with MHSCT : lipid order (% trans) at 33.4°C increased from 40 (1) to 54 (2), cooperativity decreased from 7.9 (0.4) to 5.4 (0.3), the phase transition temperature (C) increased from 30.3 (0.4) to 34.2 (0.9), and the magnitude of the phase transition (cm) increased from 4.0 (0.1) to 4.7 (0.5) (standard error of the mean). CONCLUSIONS: Conformational and thermodynamic differences were observed between Mc and MHSCT. The changes observed in the lipid conformation of meibum from patients receiving HSCTs suggest that meibum composition changes after stem cell transplantation, and clinicians should consider treating the meibomian glands to improve the ocular surface.

Protecting activity of desiccated enzymes.

Protein-based biological drugs and many industrial enzymes are unstable, making them prohibitively expensive. Some can be stabilized by formulation with excipients, but most still require low temperature storage. In search of new, more robust excipients, we turned to the tardigrade, a microscopic animal that synthesizes cytosolic abundant heat soluble (CAHS) proteins to protect its cellular components during desiccation. We find that CAHS proteins protect the test enzymes lactate dehydrogenase and lipoprotein lipase against desiccation-, freezing-, and lyophilization-induced deactivation. Our data also show that a variety of globular and disordered protein controls, with no known link to desiccation tolerance, protect our test enzymes. Protection of lactate dehydrogenase correlates, albeit imperfectly, with the charge density of the protein additive, suggesting an approach to tune protection by modifying charge. Our results support the potential use of CAHS proteins as stabilizing excipients in formulations and suggest that other proteins may have similar potential.