TCR Retrogenic Mice as a Model To Map Self-Tolerance Mechanisms to the Cancer Mucosa Antigen GUCY2C.

Characterizing self-tolerance mechanisms and their failure is critical to understand immune homeostasis, cancer immunity, and autoimmunity. However, examination of self-tolerance mechanisms has relied primarily on transgenic mice expressing TCRs targeting well-characterized, but nonphysiologic, model Ags, such as OVA and hemagglutinin. Identifying TCRs directed against bona fide self-antigens is made difficult by the extraordinary diversity of TCRs and the low prevalence of Ag-specific clones (<10-100 naive cells per organism), limiting dissection of tolerance mechanisms restricting immunity to self-proteins. In this study, we isolated and characterized TCRs recognizing the intestinal epithelial cell receptor and colorectal cancer Ag GUCY2C to establish a model to study self-antigen-specific tolerance mechanisms. GUCY2C-specific CD4+ effector T cells were isolated from immunized, nontolerant Gucy2c -/- mice. Next-generation sequencing identified GUCY2C-specific TCRs, which were engineered into CD4+ T cells in vitro to confirm TCR recognition of GUCY2C. Further, the generation of "retrogenic" mice by reconstitution with TCR-transduced hematopoietic stem cells resulted in normal CD4+ T cell development, responsiveness to immunization, and GUCY2C-induced tolerance in recipient mice, recapitulating observations in conventional models. This retrogenic model can be employed to define self-tolerance mechanisms restricting T and B cell responses to GUCY2C to optimize colorectal cancer immunotherapy without autoimmunity.

A novel multiple-stage antimalarial agent that inhibits protein synthesis.

There is an urgent need for new drugs to treat malaria, with broad therapeutic potential and novel modes of action, to widen the scope of treatment and to overcome emerging drug resistance. Here we describe the discovery of DDD107498, a compound with a potent and novel spectrum of antimalarial activity against multiple life-cycle stages of the Plasmodium parasite, with good pharmacokinetic properties and an acceptable safety profile. DDD107498 demonstrates potential to address a variety of clinical needs, including single-dose treatment, transmission blocking and chemoprotection. DDD107498 was developed from a screening programme against blood-stage malaria parasites; its molecular target has been identified as translation elongation factor 2 (eEF2), which is responsible for the GTP-dependent translocation of the ribosome along messenger RNA, and is essential for protein synthesis. This discovery of eEF2 as a viable antimalarial drug target opens up new possibilities for drug discovery.

One-Health: a Safe, Efficient, Dual-Use Vaccine for Humans and Animals against Middle East Respiratory Syndrome Coronavirus and Rabies Virus.

Middle East respiratory syndrome coronavirus (MERS-CoV) emerged in 2012 and is a highly pathogenic respiratory virus. There are no treatment options against MERS-CoV for humans or animals, and there are no large-scale clinical trials for therapies against MERS-CoV. To address this need, we developed an inactivated rabies virus (RABV) that contains the MERS-CoV spike (S) protein expressed on its surface. Our initial recombinant vaccine, BNSP333-S, expresses a full-length wild-type MERS-CoV S protein; however, it showed significantly reduced viral titers compared to those of the parental RABV strain and only low-level incorporation of full-length MERS-CoV S into RABV particles. Therefore, we developed a RABV-MERS vector that contained the MERS-CoV S1 domain of the MERS-CoV S protein fused to the RABV G protein C terminus (BNSP333-S1). BNSP333-S1 grew to titers similar to those of the parental vaccine vector BNSP333, and the RABV G-MERS-CoV S1 fusion protein was efficiently expressed and incorporated into RABV particles. When we vaccinated mice, chemically inactivated BNSP333-S1 induced high-titer neutralizing antibodies. Next, we challenged both vaccinated mice and control mice with MERS-CoV after adenovirus transduction of the human dipeptidyl peptidase 4 (hDPP4) receptor and then analyzed the ability of mice to control MERS-CoV infection. Our results demonstrated that vaccinated mice were fully protected from the MERS-CoV challenge, as indicated by the significantly lower MERS-CoV titers and MERS-CoV and mRNA levels in challenged mice than those in unvaccinated controls. These data establish that an inactivated RABV-MERS S-based vaccine may be effective for use in animals and humans in areas where MERS-CoV is endemic. IMPORTANCE: Rabies virus-based vectors have been proven to be efficient dual vaccines against rabies and emergent infectious diseases such as Ebola virus. Here we show that inactivated rabies virus particles containing the MERS-CoV S1 protein induce potent immune responses against MERS-CoV and RABV. This novel vaccine is easy to produce and may be useful to protect target animals, such as camels, as well as humans from deadly MERS-CoV and RABV infections. Our results indicate that this vaccine approach can prevent disease, and the RABV-based vaccine platform may be a valuable tool for timely vaccine development against emerging infectious diseases.

Psychiatry in American Medical Education: The Case of Harvard's Medical School, 1900-1945.

As American psychiatrists moved from the asylum to the private clinic during the early twentieth century, psychiatry acquired a growing presence within medical school curricula. This shift in disciplinary status took place at a time when medical education itself was experiencing a period of reform. By examining medical school registers at Harvard University, records from the Dean's office of Harvard's medical school, and oral histories, this paper examines the rise in prominence of psychiatry in medical education. Three builders of Harvard psychiatry - Elmer E. Southard, C. Macfie Campbell, and Harry C. Solomon - simultaneously sought to mark territory for psychiatry and its relevance. In doing so, they capitalized on three related elements: the fluidity that existed between psychiatry and neurology, the new venues whereby medical students gained training in psychiatry, and the broader role of patrons, professional associations, and certification boards, which sought to expand psychiatry's influence in the social and cultural life of twentieth-century America.

An Evidence-based Approach to the Medical Management of Fibroids: A Systematic Review.

Fibroids are the most common tumor of the female reproductive tract, but approved medical treatments are limited. Patients demand uterine-sparing treatments which preserve fertility and avoid surgery. We systematically reviewed PubMed and Cochrane databases from January 1985 to November 2015 for evidence-based medical therapies for fibroids in the context of disease prevention, treatment of early disease, treatment of symptomatic disease, and preoperative management. We identified 2182 studies, of which 52 studies met inclusion and exclusion criteria. Published data affirm the efficacy of multiple agents, which are promising avenues for the development of medical alternatives to surgery.

Split tolerance permits safe Ad5-GUCY2C-PADRE vaccine-induced T-cell responses in colon cancer patients.

BACKGROUND: The colorectal cancer antigen GUCY2C exhibits unique split tolerance, evoking antigen-specific CD8+, but not CD4+, T-cell responses that deliver anti-tumor immunity without autoimmunity in mice. Here, the cancer vaccine Ad5-GUCY2C-PADRE was evaluated in a first-in-man phase I clinical study of patients with early-stage colorectal cancer to assess its safety and immunological efficacy. METHODS: Ten patients with surgically-resected stage I or stage II (pN0) colon cancer received a single intramuscular injection of 1011 viral particles (vp) of Ad5-GUCY2C-PADRE. Safety assessment and immunomonitoring were carried out for 6 months following immunization. This trial employed continual monitoring of both efficacy and toxicity of subjects as joint primary outcomes. RESULTS: All patients receiving Ad5-GUCY2C-PADRE completed the study and none developed adverse events greater than grade 1. Antibody responses to GUCY2C were detected in 10% of patients, while 40% exhibited GUCY2C-specific T-cell responses. GUCY2C-specific responses were exclusively CD8+ cytotoxic T cells, mimicking pre-clinical studies in mice in which GUCY2C-specific CD4+ T cells are eliminated by self-tolerance, while CD8+ T cells escape tolerance and mediate antitumor immunity. Moreover, pre-existing neutralizing antibodies (NAbs) to the Ad5 vector were associated with poor vaccine-induced responses, suggesting that Ad5 NAbs oppose GUCY2C immune responses to the vaccine in patients and supported by mouse studies. CONCLUSIONS: Split tolerance to GUCY2C in cancer patients can be exploited to safely generate antigen-specific cytotoxic CD8+, but not autoimmune CD4+, T cells by Ad5-GUCY2C-PADRE in the absence of pre-existing NAbs to the viral vector. TRIAL REGISTRATION: This trial (NCT01972737) was registered at ClinicalTrials.gov on October 30th, 2013. https://clinicaltrials.gov/ct2/show/NCT01972737.

Human GUCY2C-Targeted Chimeric Antigen Receptor (CAR)-Expressing T Cells Eliminate Colorectal Cancer Metastases.

One major hurdle to the success of adoptive T-cell therapy is the identification of antigens that permit effective targeting of tumors in the absence of toxicities to essential organs. Previous work has demonstrated that T cells engineered to express chimeric antigen receptors (CAR-T cells) targeting the murine homolog of the colorectal cancer antigen GUCY2C treat established colorectal cancer metastases, without toxicity to the normal GUCY2C-expressing intestinal epithelium, reflecting structural compartmentalization of endogenous GUCY2C to apical membranes comprising the intestinal lumen. Here, we examined the utility of a human-specific, GUCY2C-directed single-chain variable fragment as the basis for a CAR construct targeting human GUCY2C-expressing metastases. Human GUCY2C-targeted murine CAR-T cells promoted antigen-dependent T-cell activation quantified by activation marker upregulation, cytokine production, and killing of GUCY2C-expressing, but not GUCY2C-deficient, cancer cells in vitro GUCY2C CAR-T cells provided long-term protection against lung metastases of murine colorectal cancer cells engineered to express human GUCY2C in a syngeneic mouse model. GUCY2C murine CAR-T cells recognized and killed human colorectal cancer cells endogenously expressing GUCY2C, providing durable survival in a human xenograft model in immunodeficient mice. Thus, we have identified a human GUCY2C-specific CAR-T cell therapy approach that may be developed for the treatment of GUCY2C-expressing metastatic colorectal cancer. Cancer Immunol Res; 6(5); 509-16. ©2018 AACR.

Between the Mind Twist and the Brain Spot. The Material Dimensions of Psychopathology in the Work of Elmer Ernest Southard .

This paper explores the material and visual practices that defined studies of psychopathology in early twentieth-century American medicine, through a close look at the work of neuropathologist Elmer E. Southard (1876­1920). As a discipline sitting at the intersection between laboratory and clinical practice, neuropathology has received little attention from historians of the brain sciences. Unlike the neurologist, who was interested in treating patients and saving lives, the neuropathologist often encountered patients following death, and studied the brain for signs of pathology during autopsy. Trained in a German tradition of laboratory pathology, Southard has been cast as a somaticist with respect to psychopathology. By examining Southard's medical and philosophical writings, I present a more nuanced analysis of the role of brain pathology in Southard's vision of disease etiology, his views on the foundations of psychiatry, as well as a more vivid picture of the sorts of material practices that defined the work of the neuropathologist.

The swinging pendulum of cancer immunotherapy personalization.

Cancer immunotherapy has long offered the promise of producing cancer treatments that are more effective and less toxic than traditional chemotherapy and radiotherapy. That potential has only begun to be realized in the last 5 years with the first US FDA-approved cancer vaccine (sipuleucel-T), checkpoint inhibitors and adoptive cell therapy. While these therapies have been remarkably more effective than previous cancer immunotherapeutics, they are often limited by their inherently personalized nature. Indeed, each patient's immune system and cancer are unique, limiting the scalability and generalizability of new approaches. However, emerging solutions may overcome these limitations, producing 'off-the-shelf' cancer immunotherapies that transform patient outcomes.

Antimalarial efficacy of MMV390048, an inhibitor of Plasmodium phosphatidylinositol 4-kinase.

As part of the global effort toward malaria eradication, phenotypic whole-cell screening revealed the 2-aminopyridine class of small molecules as a good starting point to develop new antimalarial drugs. Stemming from this series, we found that the derivative, MMV390048, lacked cross-resistance with current drugs used to treat malaria. This compound was efficacious against all Plasmodium life cycle stages, apart from late hypnozoites in the liver. Efficacy was shown in the humanized Plasmodium falciparum mouse model, and modest reductions in mouse-to-mouse transmission were achieved in the Plasmodium berghei mouse model. Experiments in monkeys revealed the ability of MMV390048 to be used for full chemoprotection. Although MMV390048 was not able to eliminate liver hypnozoites, it delayed relapse in a Plasmodium cynomolgi monkey model. Both genomic and chemoproteomic studies identified a kinase of the Plasmodium parasite, phosphatidylinositol 4-kinase, as the molecular target of MMV390048. The ability of MMV390048 to block all life cycle stages of the malaria parasite suggests that this compound should be further developed and may contribute to malaria control and eradication as part of a single-dose combination treatment.

Are resident work-hour limitations beneficial to the trauma profession?

In July 2003, work-hour restrictions were implemented by the Accreditation Council for Graduate Medical Education (ACGME) to limit resident duty hours. Attending surgeon work-hours have not been similarly reduced, and many trauma services have added emergency general surgery responsibilities. We hypothesized that trauma attending/resident work-hour disparity may disincentivize residents from selecting trauma careers and that trauma directors would view ACGME regulations negatively. We conducted a 6-month study of resident and in-house trauma attending self-reported hours at a level I trauma center and sent a questionnaire to 172 national level I trauma directors (TDs) regarding work-hours restrictions. TD survey response rate was 48 per cent; 100 per cent of 15 residents and 6 trauma faculty completed work-hour logs. Attending mean hours (87.1/ wk), monthly calls (5), and shifts > 30 hours exceeded that of all resident groups. Case volume was similar. Residents viewed their lifestyle more favorably than the lifestyle of the trauma attending (Likert score 3.6 +/- 0.5 vs Likert score 2.5 +/- 0.8, P = 0.0003). Seventy-one per cent cited attending work hours and lifestyle as a reason not to pursue a trauma career. Nationally, 80 per cent of trauma surgeons cover emergency general surgery; 40 per cent work greater than 80 hours weekly, compared with < 1 per cent of surgical trainees (P < 0.0001). Most TDs feel that residents do not spend more time reading (89%) or operating (96%); 68 per cent feel patient care has suffered as a result of duty-hours restrictions. Seventy-one per cent feel residents will not select trauma surgery as a career as a result of changes in duty hours. Perceived trauma attending/ resident work-hour disparity may disincentive trainees from trauma career selection. TDs view resident duty-hour restrictions negatively.

GUCY2C-directed CAR-T cells oppose colorectal cancer metastases without autoimmunity.

Adoptive T-cell therapy (ACT) is an emerging paradigm in which T cells are genetically modified to target cancer-associated antigens and eradicate tumors. However, challenges treating epithelial cancers with ACT reflect antigen targets that are not tumor-specific, permitting immune damage to normal tissues, and preclinical testing in artificial xenogeneic models, preventing prediction of toxicities in patients. In that context, mucosa-restricted antigens expressed by cancers exploit anatomical compartmentalization which shields mucosae from systemic antitumor immunity. This shielding may be amplified with ACT platforms employing antibody-based chimeric antigen receptors (CARs), which mediate MHC-independent recog-nition of antigens. GUCY2C is a cancer mucosa antigen expressed on the luminal surfaces of the intestinal mucosa in mice and humans, and universally overexpressed by colorectal tumors, suggesting its unique utility as an ACT target. T cells expressing CARs directed by a GUCY2C-specific antibody fragment recognized GUCY2C, quantified by expression of activation markers and cytokines. Further, GUCY2C CAR-T cells lysed GUCY2C-expressing, but not GUCY2C-deficient, mouse colorectal cancer cells. Moreover, GUCY2C CAR-T cells reduced tumor number and morbidity and improved survival in mice harboring GUCY2C-expressing colorectal cancer metastases. GUCY2C-directed T cell efficacy reflected CAR affinity and surface expression and was achieved without immune-mediated damage to normal tissues in syngeneic mice. These observations highlight the potential for therapeutic translation of GUCY2C-directed CAR-T cells to treat metastatic tumors, without collateral autoimmunity, in patients with metastatic colorectal cancer.

Integrating mind and brain: Warren S. McCulloch, cerebral localization, and experimental epistemology.

Recently, historians have focused on Warren S. McCulloch's role in the cybernetics movement during the 1940s and 1950s, and his contributions to the development of computer science and communication theory. What has received less attention is McCulloch's early work in neurophysiology, and its relationship to his philosophical quest for an 'experimental epistemology' - a physiological theory of knowledge. McCulloch's early laboratory work during the 1930s addressed the problem of cerebral localization: localizing aspects of behaviour in the cerebral cortex of the brain. Most of this research was done with the Dutch neurophysiologist J.G. Dusser de Barenne at Yale University. The connection between McCulloch's philosophical interests and his experimental work can be expressed as a search for a physiological a priori, an integrated mechanism of sensation.

Transcending disciplines: Scientific styles in studies of the brain in mid-twentieth century America.

Much scholarship in the history of cybernetics has focused on the far-reaching cultural dimensions of the movement. What has garnered less attention are efforts by cyberneticians such as Warren McCulloch and Norbert Wiener to transform scientific practice in an array of disciplines in the biomedical sciences, and the complex ways these efforts were received by members of traditional disciplines. In a quest for scientific unity that had a decidedly imperialistic flavour, cyberneticians sought to apply practices common in the exact sciences-mainly theoretical modeling-to problems in disciplines that were traditionally defined by highly empirical practices, such as neurophysiology and neuroanatomy. Their efforts were met with mixed, often critical responses. This paper attempts to make sense of such dynamics by exploring the notion of a scientific style and its usefulness in accounting for the contrasts in scientific practice in brain research and in cybernetics during the 1940s. Focusing on two key institutional contexts of brain research and the role of the Rockefeller and Macy Foundations in directing brain research and cybernetics, the paper argues that the conflicts between these fields were not simply about experiment vs. theory but turned more closely on the questions that defined each area and the language used to elaborate answers.

Economic burden of dermatologic adverse events induced by molecularly targeted cancer agents.

OBJECTIVE: To report the financial impact of diagnosing and treating the dermatologic toxicities (dTs) that develop in patients receiving targeted anticancer therapies. DESIGN: Single-center retrospective and prospective medical record data extraction. SETTING: Department of Dermatology, Northwestern University, Chicago, Illinois. PATIENTS: One hundred thirty-two adults who presented between November 1, 2005, and June 30, 2008, and who were diagnosed as having 1 primary cancer type and were treated with 1 molecularly targeted agent. MAIN OUTCOME MEASURE: Standard billable costs to the patient for dT-related medications, clinic visits, laboratory and diagnostic testing, and therapeutic procedures. RESULTS: The 132 patients had a median of 3 clinic visits for dT management with a median cost of $1920 per patient. Sorafenib was associated with the most costly overall median cost per patient ($2509 per patient), and imatinib was associated with the least costly overall median cost per patient ($1263 per patient). Among the 7 targeted drugs and all 10 dTs, the most costly dT (measured by cost of treatment with medications) was hand/foot skin reaction, associated with sorafenib therapy (median cost, $968 per patient) (P < .001). The second most costly dT was panitumumab-associated acneiform eruption (median cost, $933 per patient) (P < .001). CONCLUSION: The cost of diagnosis and treatment of dTs associated with targeted agents contributes to the overall economic burden of cancer care. Efforts toward the prevention of dTs may be important for decreasing the financial burden in oncology.

(Physio)logical circuits: the intellectual origins of the McCulloch-Pitts neural networks.

This article examines the intellectual and institutional factors that contributed to the collaboration of neuropsychiatrist Warren McCulloch and mathematician Walter Pitts on the logic of neural networks, which culminated in their 1943 publication, "A Logical Calculus of the Ideas Immanent in Nervous Activity." Historians and scientists alike often refer to the McCulloch-Pitts paper as a landmark event in the history of cybernetics, and fundamental to the development of cognitive science and artificial intelligence. This article seeks to bring some historical context to the McCulloch-Pitts collaboration itself, namely, their intellectual and scientific orientations and backgrounds, the key concepts that contributed to their paper, and the institutional context in which their collaboration was made. Although they were almost a generation apart and had dissimilar scientific backgrounds, McCulloch and Pitts had similar intellectual concerns, simultaneously motivated by issues in philosophy, neurology, and mathematics. This article demonstrates how these issues converged and found resonance in their model of neural networks. By examining the intellectual backgrounds of McCulloch and Pitts as individuals, it will be shown that besides being an important event in the history of cybernetics proper, the McCulloch-Pitts collaboration was an important result of early twentieth-century efforts to apply mathematics to neurological phenomena.