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Pancreatic ductal adenocarcinoma (PDA) is a lethal malignancy resistant to therapies, including immune-checkpoint blockade. We investigated two distinct strategies to modulate tumor-associated macrophages (TAM) to enhance cellular therapy targeting mesothelin in an autochthonous PDA mouse model. Administration of an antibody to colony-stimulating factor (anti-Csf1R) depleted Ly6Clow protumorigenic TAMs and significantly enhanced endogenous T-cell intratumoral accumulation. Despite increasing the number of endogenous T cells at the tumor site, as previously reported, TAM depletion had only minimal impact on intratumoral accumulation and persistence of T cells engineered to express a murine mesothelin-specific T-cell receptor (TCR). TAM depletion interfered with the antitumor activity of the infused T cells in PDA, evidenced by reduced tumor cell apoptosis. In contrast, TAM programming with agonistic anti-CD40 increased both Ly6Chigh TAMs and the intratumoral accumulation and longevity of TCR-engineered T cells. Anti-CD40 significantly increased the frequency and number of proliferating and granzyme B+ engineered T cells, and increased tumor cell apoptosis. However, anti-CD40 failed to rescue intratumoral engineered T-cell IFNγ production. Thus, although functional modulation, rather than TAM depletion, enhanced the longevity of engineered T cells and increased tumor cell apoptosis, ultimately, anti-CD40 modulation was insufficient to rescue key effector defects in tumor-reactive T cells. This study highlights critical distinctions between how endogenous T cells that evolve in vivo, and engineered T cells with previously acquired effector activity, respond to modifications of the tumor microenvironment.

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Adoptive T cell therapy (ACT) has demonstrated activity in solid tumors but requires further optimization to become a reproducibly effective treatment. T cell receptor (TCR)-engineered T cells recognize peptides derived from intracellular and surface proteins presented in the context of MHC class I. Targeting mutated oncogenic drivers addresses some of the major obstacles of this modality, in that the antigenic epitope is: 1) tumor-specific, 2) essential for tumor survival, and 3) derived from a stably expressed protein. KRAS is the most frequently mutated gene in human cancers with alterations in codon 12 associated with poor clinical outcomes in a high proportion of colon, lung and pancreatic cancers, as well as many others. To isolate TCRs specific for the peptide derived from the KRAS G12D mutation presented in the context of HLA-A*11:01, one of the most common HLA alleles worldwide, we employed our high-throughput in vitro TCR discovery platform. CD8+ T cells were isolated from healthy donors and co-cultured with autologous antigen-presenting cells exogenously-loaded with mutant KRAS peptides encompassing the G12D mutation. The highest avidity T cells were subsequently identified and enriched by fluorescence-based cell sorting and the corresponding TCR genes isolated by single-cell sequencing and inserted into a lentiviral expression vector. TCR candidates were transduced into primary T cells and prioritized based on functional avidity and specificity (response to titrated peptide-loaded presenting cells and tumor cells endogenously expressing the KRAS G12D antigen) and cytotoxicity (in vitro tumor cell killing assays). X-Scan studies, in which each residue of the reference KRAS G12D peptide was systematically substituted by all other amino acids, revealed a highly restrictive TCR recognition motif, suggesting limited risk of promiscuous off-target activation. We engineered both CD4+ and CD8+ T cells to lentivirally-express candidate TCRs in addition to the genes encoding the CD8αβ co-receptor to enhance TCR-HLA class I avidity in CD4+ T cells, with the aim of creating a coordinated CD4 and CD8 T cell response to the same tumor target to promote increased T cell activity and persistence while minimizing T cell exhaustion. Transduced CD4+ T cells were functional and could be demonstrated to provide help to CD8+ T cells, supporting tumor elimination in several experimental models. In summary, we report a TCR gene therapy approach targeting mutant KRAS G12D-containing peptides with a coordinated CD4 and CD8 T cell response that has a promising efficacy and safety profile. Our work to date supports the planned clinical development of this novel TCR-engineered T cell therapy for treating KRAS-mutant solid tumors. Citation Format: Cheryl Black, James Parsons, Anthony Thomas, Allison P. Drain, Santosh Narayan, Joshua Francis, Xingyue He, Ankit Gupta, Jessica Webb, Thomas M. Schmitt, Philip D. Greenberg, Gary Shapiro, Loïc Vincent. Preclinical development of safe and effective T cell receptors specific for mutant KRAS G12D peptide [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 1127.

Multiple Sclerosis is a chronic autoimmune neurodegenerative disorder of the central nervous system (CNS) that is characterized by inflammation, demyelination, and axonal injury leading to permeant disability. In the early stage of MS, inflammation is the primary driver of the disease progression. There remains an unmet need to develop high efficacy therapies with superior safety profiles to prevent the inflammation processes leading to disability. Herein, we describe the discovery of BIIB091, a structurally distinct orthosteric ATP competitive, reversible inhibitor that binds the BTK protein in a DFG-in confirmation designed to sequester Tyr-551, an important phosphorylation site on BTK, into an inactive conformation with excellent affinity. Preclinical studies demonstrated BIB091 to be a high potency molecule with good drug-like properties and a safety/tolerability profile suitable for clinical development as a highly selective, reversible BTKi for treating autoimmune diseases such as MS.

Pancreatic ductal adenocarcinoma (PDA) is a lethal malignancy resistant to therapies, including immune-checkpoint blockade. We investigated two distinct strategies to modulate tumor-associated macrophages (TAM) to enhance cellular therapy targeting mesothelin in an autochthonous PDA mouse model. Administration of an antibody to colony-stimulating factor (anti-Csf1R) depleted Ly6Clow protumorigenic TAMs and significantly enhanced endogenous T-cell intratumoral accumulation. Despite increasing the number of endogenous T cells at the tumor site, as previously reported, TAM depletion had only minimal impact on intratumoral accumulation and persistence of T cells engineered to express a murine mesothelin-specific T-cell receptor (TCR). TAM depletion interfered with the antitumor activity of the infused T cells in PDA, evidenced by reduced tumor cell apoptosis. In contrast, TAM programming with agonistic anti-CD40 increased both Ly6Chigh TAMs and the intratumoral accumulation and longevity of TCR-engineered T cells. Anti-CD40 significantly increased the frequency and number of proliferating and granzyme B+ engineered T cells, and increased tumor cell apoptosis. However, anti-CD40 failed to rescue intratumoral engineered T-cell IFNγ production. Thus, although functional modulation, rather than TAM depletion, enhanced the longevity of engineered T cells and increased tumor cell apoptosis, ultimately, anti-CD40 modulation was insufficient to rescue key effector defects in tumor-reactive T cells. This study highlights critical distinctions between how endogenous T cells that evolve in vivo, and engineered T cells with previously acquired effector activity, respond to modifications of the tumor microenvironment.

Susan Glaspell’s Poetics and Politics of Rebellion is an expansive critical study of Glaspell’s dramatic oeuvre as a drama of rebellion, treating of the various rebellion(s) staged by Glaspell and the relationship between Glaspell’s rebellious content (politics) and her innovative form (poetics). This study strengthens Glaspell’s reputation as a feminist, modernist, and profoundly political American playwright.

Objective: The convective flow of CSF plays a crucial role for CNS to clear endogenous and xenobiotic substances. The objective of this study is to investigate the impact of modifying CSF flow with acetazolamide and arginine vasopressin (AVP) on the CNS clearance of ibuprofen and acetaminophen. Results: Microdialysis studies indicated that acetaminophen AUC ratio (Kp,uu) between brain ISF and unbound plasma increased from 0.40 ± 0.14 in the vehicle control group to 0.60 ± 0.27 in the acetazolamide treated group (P < 0.05). Conversely, acetaminophen’s steady-state ISFC to unbound_plasmaC ratio (Kp,uu) decreased from 0.44 ± 0.08 to 0.36 ± 0.07 upon IV infusion of 0.3 µg/hr AVP (P < 0.05). Using CSF concentration as a surrogate of unbound brain drug concentration, AVP treatment reduced the CSFAUC0-5hr/unbound_plasmaAUC0-5hr ratio from 1.63 to 0.85 for acetaminophen, and the CSFC4hr / unbound_plasmaC4hr ratio decreased from 0.91 ± 0.27 to 0.54 ± 0.12 for ibuprofen (P < 0.05). Conclusion: We have demonstrated that acetazolamide decreases the CNS clearance of acetaminophen, while AVP increases the CNS clearance of both acetaminophen and ibuprofen. Such changes are caused by altering the CSF production rates.

Kitchen Sink Realisms examines both iconic and now-obscure works of American drama focused on domesticity and domestic labour. Chansky’s research is both extensive and sharply focused, and the study is remarkable for its breadth and its depth, its originality, and the persuasiveness of its arguments.

Edna St. Vincent Millay was a poet, playwright and free-spirited bohéme who epitomized the aesthetically and sexually adventurous ‘new woman’ of the early twentieth century. Born in Rockland, Maine, and raised by an unconventional mother who encouraged her intellectual and artistic pursuits, Millay garnered national attention at the age of nineteen with her lyric poem Renascence (1912). Millay’s innovative poetry, for which she received a Pulitzer Prize in 1923, integrated conventional forms with unconventional content that celebrated sexual liberation and female autonomy.

Founded in Provincetown, Massachusetts in 1915 and transplanted to Greenwich Village in 1916, the Provincetown Players was one of the most influential theatrical organizations in American theater history. Their membership was a veritable who’s who of the era’s leading political and cultural revolutionaries, including its spiritus rector, socialist writer George Cram (Jig) Cook; postimpressionist artists William and Marguerite Zorach and Bror Nordfeldt; labor journalists John Reed and Mary Heaton Vorse; modernist poets Wallace Stevens, Alfred Kreymborg, Mina Loy, and Edna St Vincent Millay; New Stagecraft pioneers Robert Edmond Jones and Cleon Throckmorton; and more than fifty playwrights whose dramaturgical innovations defied contemporary critical description. In addition to formal experimentation, Provincetown playwrights were noted for their frank treatment of such topical issues as racial, ethnic, and religious Otherness, class conflict, war, and changing gender and sexual mores. Their plays manifested the most current trends in the era’s intellectual discourse: Freudian and Jungian psychology; Nietzschean challenges to traditional morality; Havelock Ellis’s and Ellen Keys’ ideas on egalitarian sexual and marital relationships; the social theories of Karl Marx and Edward Carpenter; the feminism of Emma Goldman, Margaret Sanger, and Crystal Eastman; and the parenting techniques of Maria Montessori.