Nutritional Management of Infants With a Giant Ruptured Omphalocele: Clinical Observations From a Single Institution Case Series.

The nutritional requirements of neonates with congenital abdominal wall defects (AWDs) remain poorly described. In particular, there is a lack of literature on the calorie, protein, and micronutrient needs of those with AWD. Nutritional therapy is a cornerstone of care in patients with burns due to the metabolic consequences of injury to the epithelial layer. Similarly, children with AWD may require specialized nutritional plans to support their growth and wound healing. This case series supports the theory that patients with ruptured omphaloceles may require higher calorie, protein, and micronutrient provisions in comparison to patients with intact omphaloceles, due to increased metabolic demand to support wound healing and skin epithelialization.

Oral Feeding in Infants After Congenital Diaphragmatic Hernia Repair While on Non-invasive Positive Pressure Ventilation: The Impact of a Dysphagia Provider-Led Protocol.

Infants with congenital diaphragmatic hernia (CDH) who require non-invasive positive pressure ventilation or high flow nasal cannula are at risk for aspiration and delayed initiation of oral feeding. We developed a dysphagia provider-led protocol that involved early consultation with an occupational therapist or speech/language pathologist and modified barium swallow study (MBSS) to assess for readiness for oral feeding initiation/advancement on non-invasive positive pressure ventilation. The objective of this study was to retrospectively compare this intervention cohort to a historical control cohort to evaluate the protocol's impact on the time to initiate oral feeding. We describe the development and implementation of the protocol, the MBSS findings of the intervention cohort, and compared the control (n = 64) and intervention (n = 37) cohorts using Fischer's exact test and Mann-Whitney test. We found that both cohorts had similar prenatal and neonatal characteristics including age at extubation. Significantly more infants in the intervention cohort were on non-invasive positive pressure ventilation or high flow nasal cannula at the time of oral feeding initiation (84% vs. 28%, p < 0.0001). None of the control cohort infants underwent MBSS while on respiratory support. Of the intervention cohort, 15 infants underwent a MBSS while on non-invasive positive pressure ventilation; 6 had no evidence of laryngeal penetration and/or aspiration during swallowing. Infants in the control cohort initiated oral feeds significantly sooner after extubation (6 versus 11 days, p = 0.001) and attained full oral feeds earlier (20 days versus 28 days, p = 0.02) than the intervention group. There was no difference in the rate of gastrostomy tube placement (38%). Appropriate monitoring by a dysphagia provider and evaluation with clinical and radiological means are crucial to determine the safety of initiating oral feeding in term infants with CDH. Continued surveillance is needed to determine the long-term impact on oral feeding progression in this population.

Factors and Growth Trends Associated With the Need for Gastrostomy Tube in Neonates With Congenital Diaphragmatic Hernia.

OBJECTIVES: A third of infants with congenital diaphragmatic hernia (CDH) require a gastrostomy tube (GT) for nutritional support. We compared CDH infants who are GT-dependent to those able to meet their nutritional needs orally, to identify factors associated with requiring a GT and evaluate their long-term growth. METHODS: Patients with CDH repaired at a single institution between 2012 and 2020 were included. Charts were retrospectively reviewed for demographic, surgical, and post-operative details. Mann-Whitney test and Fischer exact test were performed to compare GT-dependent neonates (n = 38, experimental) with orally fed neonates (n = 63, control). Significance was set at <0.05. RESULTS: Thirty-eight percent received a GT (median 67 days, interquartile range [IQR] 50-88). GT-dependent neonates were significantly more likely to have a lower lung-to-head ratio (median 1.2, IQR 0.9-1.4, vs 1.6, IQR 1.3-2.0, IQR P < 0.0001), undergone patch or flap repair (79% vs 33%, P < 0.0001), and been hospitalized longer (median 47, IQR 24-75 vs 28 days, P < 0.0001). Fourteen of 38 had their GT removed (median 26 months, IQR 14-36). GT-dependent neonates initiated oral feeds (calculated as time since extubation) later (median 21, IQR 8-26, vs 8 days, IQR 4-13, P = 0.006). Height-for-age z scores remained stable after GT removal, while weight-for-age z scores dropped initially and began improving a year later. CONCLUSIONS: The need for a gastrostomy for nutritional support is associated with more severe CDH. Over a third of patients no longer needed a GT at a median of 26 months. Linear growth generally remains stable after removal. These results may help counsel parents regarding nutritional expectations.

Oncolytic Newcastle disease virus activation of the innate immune response and priming of antitumor adaptive responses in vitro.

Oncolytic virus (OV) therapy is an emerging approach with the potential to redefine treatment options across a range of cancer indications and in patients who remain resistant to existing standards of care, including immuno-oncology (IO) drugs. MEDI5395, a recombinant Newcastle disease virus (NDV), engineered to express granulocyte-macrophage colony-stimulating factor (GM-CSF), exhibits potent oncolytic activity. It was hypothesized that activation of immune cells by MEDI5395, coupled with its oncolytic activity, would enhance the priming of antitumor immunity. Using MEDI5395 and recombinant NDVs encoding fluorescent reporter genes, we demonstrated preferential virus uptake and non-productive infection in myeloid cells, including monocytes, macrophages, and dendritic cells (DCs). Infection resulted in immune-cell activation, with upregulation of cell surface activation markers (e.g., CD80, PD-L1, HLA-DR) and secretion of proinflammatory cytokines (IFN-α2a, IL-6, IL-8, TNF-α). Interestingly, in vitro M2-polarized macrophages were more permissive to virus infection than were M1-polarized macrophages. In a co-culture system, infected myeloid cells were effective virus vectors and mediated the transfer of infectious NDV particles to tumor cells, resulting in cell death. Furthermore, NDV-infected DCs stimulated greater proliferation of allogeneic T cells than uninfected DCs. Antigens released after NDV-induced tumor cell lysis were cross-presented by DCs and drove activation of tumor antigen-specific autologous T cells. MEDI5395 therefore exhibited potent immunostimulatory activity and an ability to enhance antigen-specific T-cell priming. This, coupled with its tumor-selective oncolytic capacity, underscores the promise of MEDI5395 as a multimodal therapeutic, with potential to both enhance current responding patient populations and elicit de novo responses in resistant patients.

Multifaceted Sexual Desire and Hormonal Associations: Accounting for Social Location, Relationship Status, and Desire Target.

Sexual desire is typically measured as a unitary erotic phenomenon and is often assumed by biological and biomedical researchers, as well as the lay public, to be directly connected to physiological parameters like testosterone (T). In the present study, we empirically examined how conceptualizing sexual desire as multifaceted might clarify associations with T and contextual variables. To do so, we used the Sexual Desire Questionnaire (DESQ), which assesses multifaceted dyadic sexual desire, to explore how contextual variables such as social location, relationship status, and desire target (e.g., partner vs. stranger) might be meaningful for reports of sexual desire and associated hormonal correlations. We focused on women (N = 198), because sexual desire and testosterone are generally unlinked in healthy men. Participants imagined a partner or stranger while answering the 65 DESQ items and provided a saliva sample for hormone assay. Analyses showed that the DESQ factored differently for the current sample than in previous research, highlighting how sexual desire can be constructed differently across different populations. We also found that, for the Intimacy, Eroticism, and Partner Focus factors, mean scores were higher when the desire target was a partner relative to a stranger for participants in a relationship, but equally high between partner versus stranger target for single participants. DESQ items resolved into meaningful hormonal desire components, such that high endorsement of Fantasy Experience was linked to higher T, and higher cortisol was linked with lower endorsement of the Intimacy factor. We argue that conceptualizing desire as multifaceted and contextualized when assessing hormonal links-or questions in general about desire-can clarify some of its complexities and lead to new research avenues.

Sexual Desire in Sexual Minority and Majority Women and Men: The Multifaceted Sexual Desire Questionnaire.

Sexual desire is increasingly understood to be multifaceted and not solely erotically oriented, but measures are still generally unitary and eroticism-focused. Our goals in this article were to explore the multifaceted nature of sexual desire and develop a measure to do so, and to determine how multifaceted sexual desire might be related to gender/sex and sexual orientation/identity. In the development phase, we generated items to form the 65-item Sexual Desire Questionnaire (DESQ). Next, the DESQ was administered to 609 women, 705 men, and 39 non-binary identified participants. Results showed that the DESQ demonstrated high reliability and validity, and that sexual desire was neither unitary nor entirely erotic, but instead was remarkably multifaceted. We also found that multifaceted sexual desire was in part related to social location variables such as gender/sex and sexual orientation/identity. We propose the DESQ as a measure of multifaceted sexual desire that can be used to compare factor themes, total scores, and scores across individual items in diverse groups that take social context into account. Results are discussed in light of how social location variables should be considered when making generalizations about sexual desire, and how conceptualizations of desire as multifaceted may provide important insights.

Critical roles of Bcl11b in T-cell development and maintenance of T-cell identity.

T-cell development primarily occurs in the thymus and involves in the interactions of many important transcription factors. Until recently, no single transcription factor has been identified to be essential for T-cell lineage commitment or maintenance of T-cell identity. Recent studies have now identified the zinc finger transcription factor Bcl11b to be essential for T-cell development and for maintenance of T-cell identity. Remarkably, T cells acquire NK cell properties upon Bcl11b deletion. These reprogrammed cells have unique properties in proliferation, cytokine dependency and killing target cells, and may therefore provide a new cell source for some cell-based therapies.

Mediation of antitumor activity by AZD4820 oncolytic vaccinia virus encoding IL-12.

Oncolytic viruses are engineered to selectively kill tumor cells and have demonstrated promising results in early-phase clinical trials. To further modulate the innate and adaptive immune system, we generated AZD4820, a vaccinia virus engineered to express interleukin-12 (IL-12), a potent cytokine involved in the activation of natural killer (NK) and T cells and the reprogramming of the tumor immune microenvironment. Testing in cultured human tumor cell lines demonstrated broad in vitro oncolytic activity and IL-12 transgene expression. A surrogate virus expressing murine IL-12 demonstrated antitumor activity in both MC38 and CT26 mouse syngeneic tumor models that responded poorly to immune checkpoint inhibition. In both models, AZD4820 significantly upregulated interferon-gamma (IFN-γ) relative to control mice treated with oncolytic vaccinia virus (VACV)-luciferase. In the CT26 study, 6 of 10 mice had a complete response after treatment with AZD4820 murine surrogate, whereas control VACV-luciferase-treated mice had 0 of 10 complete responders. AZD4820 treatment combined with anti-PD-L1 blocking antibody augmented tumor-specific T cell immunity relative to monotherapies. These findings suggest that vaccinia virus delivery of IL-12, combined with immune checkpoint blockade, elicits antitumor immunity in tumors that respond poorly to immune checkpoint inhibitors.

New views on natural killer cell-based immunotherapy for melanoma treatment.

Natural killer (NK) cell-based immunotherapies treat hematopoietic malignancies, but are less effective against solid tumors. Here, we review recent data on NK cell recognition of melanoma at various stages of the disease and propose a combinatorial strategy to exploit fully the potential of NK cells. Depending on the stage of melanoma progression, NK cell-based therapies could be combined with pharmacological and T cell-based immunotherapies, to: (i) prevent lymph node metastases by redistributing cytotoxic NK cells; (ii) boost NK cell activity using chemotherapy to upregulate activating ligands on tumor cells; and (iii) target visceral metastases by transfer of autologous or allogeneic NK cells.

Advancing a grounded theory of parental support in competitive girls' golf.

Parental support in youth sport has been associated with positive athlete outcomes, such as sport enjoyment and continued participation. Although research has demonstrated the significant and influential role parents fulfil in the youth sport context, there remains a dearth of theoretical frameworks detailing parental support in youth sport and an absence of empirical research examining parental support across athlete development stages and sports. The present study sought to examine athletes' perceptions of parental support, with a view to advancing a grounded theory of parental support in youth golf. Fourteen online synchronous focus groups were conducted with an international sample (Australia, Canada, England, Finland, Ireland, New Zealand, Scotland) of 61 girls, in the specialising (n = 27) and investment stages (n = 34) of athlete development. Data were analysed in three phases: open-coding, axial coding, and theoretical integration. The substantive grounded theory is constructed on the core category of 'Individual Parental Support Preferences'. This core category is underpinned by four sub-categories of parental support which were evident across development stages: instrumental, informational, emotional, and autonomy support, and is influenced by a host of athlete (e.g., athletes' performance), parent (e.g., parents' knowledge), and contextual characteristics (e.g., location). Unconditional parental support is an important aspect of emotional support, however the concept of adopting a person-first approach to sport parenting is novel. These results provide a rich and novel insight of parental support in girls' golf, advancing a grounded theoretical understanding of parental support mechanisms in a youth sport context.

Emergency Medicine Obstetrics and Gynecology: A Case-Based Curriculum for Residents.

Introduction: Millions of patients present to US emergency departments every year with OB/GYN concerns. Emergency medicine trainees must be adequately prepared to care for this population, regardless of how commonly they appear in the training environment. We used active learning and gamification principles in this curriculum to increase learner engagement and participation in the material. Methods: We chose OB/GYN topics based on review of Tintinalli's OB/GYN content and the American Board of Emergency Medicine's Model of Clinical Practice. Each session comprised a case-based lecture and review questions using the game-based Kahoot! online software. Pre- and postcurriculum surveys assessed residents' confidence in caring for emergent OB/GYN pathologies on a 5-point Likert scale. We designed survey questions assessing the first level of Kirkpatrick's levels of training evaluation; these questions were reviewed and revised by the department's Medical Education Scholarship Committee for validity. Results: A mean of 18 residents attended each session. Seventy-six percent of residents (26 of 34) completed the precurriculum survey, 67% (23 of 34) completed the postcurriculum survey, and 44% (15 of 34) completed both. For all respondents, mean reported confidence with curriculum topics increased from 3.5 to 4.0 (p < .05). For residents completing both surveys, confidence increased from 3.4 to 4.0 (p < .01). Discussion: Application of this curriculum significantly improved learner confidence in targeted OB/GYN topics. Future directions could include evaluating curricular impact at higher levels in the Kirkpatrick model, extending sessions to include more time for interaction, and adding suggested readings.

Global dataset of soil organic carbon in tidal marshes.

Tidal marshes store large amounts of organic carbon in their soils. Field data quantifying soil organic carbon (SOC) stocks provide an important resource for researchers, natural resource managers, and policy-makers working towards the protection, restoration, and valuation of these ecosystems. We collated a global dataset of tidal marsh soil organic carbon (MarSOC) from 99 studies that includes location, soil depth, site name, dry bulk density, SOC, and/or soil organic matter (SOM). The MarSOC dataset includes 17,454 data points from 2,329 unique locations, and 29 countries. We generated a general transfer function for the conversion of SOM to SOC. Using this data we estimated a median (± median absolute deviation) value of 79.2 ± 38.1 Mg SOC ha-1 in the top 30 cm and 231 ± 134 Mg SOC ha-1 in the top 1 m of tidal marsh soils globally. This data can serve as a basis for future work, and may contribute to incorporation of tidal marsh ecosystems into climate change mitigation and adaptation strategies and policies.

Multiple mechanisms contribute to impairment of type 1 interferon production during chronic lymphocytic choriomeningitis virus infection of mice.

Type 1 IFNs, innate cytokines with important effector and immunomodulatory properties, are rapidly induced in the acute phase of many virus infections; however, this is generally a transient response that is not sustained during virus persistence. To gain insight into mechanisms that can contribute to down-regulation of type 1 IFN production during virus persistence, we analyzed type 1 IFN production during acute and chronic lymphocytic choriomeningitis virus (LCMV) infection. High-level type 1 IFN production was transiently up-regulated in cells including plasmacytoid and conventional dendritic cells (DCs) following LCMV infection of mice, but LCMV persistence was associated with only low-level type 1 IFN production. Nonetheless, chronically infected mice were able to up-regulate type 1 IFN production in response to TLR3, 7, and 9 ligands, albeit less efficiently than uninfected mice. Splenic DC numbers in mice chronically infected with LCMV were decreased, and the remaining cells exhibited a reduced response to TLR stimulation. LCMV-infected cell lines efficiently up-regulated type 1 IFN production following TLR ligation and infection with a DNA virus, but exhibited a defect in type 1 IFN induction following infection with Sendai, an RNA virus. This block in type 1 IFN production by infected cells, together with abnormalities in DC numbers and functions, likely contribute to the low-level type 1 IFN production in mice chronically infected with LCMV. Impairment of type 1 IFN production may both promote virus persistence and impact on host immunocompetence. Understanding the mechanisms involved may assist in development of strategies for control of virus persistence and superinfection.

A Novel Virtual Emergency Medicine Residents-as-Teachers (RAT) Curriculum.

Audience: The Residents-as-Teachers (RAT) curriculum is designed for emergency medicine (EM) residents of all years (PGY1-4). Length of Curriculum: The curriculum is divided into three hour-long sessions. The entire curriculum can be run as a single block or can be spread out over multiple days. Introduction: The Accreditation Council of Graduate Medical Education (ACGME) and the Liaison Committee on Medical Education (LCME) both require residents to receive training in teaching medical students and junior residents. They also require opportunities for residents to participate in teaching and be assessed on their effectiveness in this role.1,2 However, the ACGME does not provide guidance or require formal curricula on molding residents into effective teachers. Many programs and institutions have incorporated RAT curricula as a solution to provide residents with the skills necessary to create an excellent educational environment for junior learners. These curricula have been embraced by many specialties, including Emergency Medicine (EM).3-6 The effectiveness of the teaching received during the clinical rotations has important long-term effects on medical students, and may impact their future career choices in medicine.6The COVID-19 pandemic has also required education institutions to vastly alter the delivery of their didactics, including moving to a virtual platform. A completely online format has many benefits that extend beyond the pandemic, such as easier access to participants (including those off-service or at remote sites), no requirement for a physical space, and easier recording of sessions. Educational Goals: To provide residents with an introduction to teaching techniques that can be utilized on-shift to facilitate an excellent educational experience for junior learners while balancing the resident's patient care responsibilities. Educational Methods: The educational strategies used in this curriculum include PowerPoint (Redmond, WA) slideshows given by a live presenter via the telecommunications platform Zoom (San Jose, CA), viewing of videos demonstrating curriculum topics, simulation-based learning through role-play, and small-group discussions including simulation debriefing. Research Methods: A survey was distributed to residents before and following the completion of the three training sessions to assess resident satisfaction with the delivery of the content and comfort with the teaching tools discussed. Suggestions on potential improvements were also assessed to inform changes to future iterations of the curriculum. Comfort regarding the included teaching tools was assessed using a five-point Likert scale. After completion of the curriculum, rotating medical students were provided with an evaluation form to assess if residents were teaching using the techniques from the course. Results: Both the pre-curriculum and post-curriculum surveys had a response rate of 61.1%. Student's t-test showed a statistically significant increase in mean resident comfort level with the teaching strategies post-curriculum (3.05 to 3.83, p < 0.01). Medical student evaluations have shown, overall, that the majority of residents are utilizing the education techniques on-shift. There were no significant differences found in medical student perception of resident use of taught skills between those who had and had not attended the sessions. However, all but one assessed skill showed higher utilization in those who had attended the correlating session. Discussion: The educational content was effective in improving the residents' comfort with the teaching strategies presented, and residents are utilizing these techniques on-shift. Through implementation, we discovered that presenting a curriculum over video conferencing required additional administrative support to help ensure efficacy of break-out groups. Based on resident feedback after the first session, multiple changes were made, including providing residents with hand-out references for use during the role-playing sessions. The success of this curriculum demonstrated the feasibility and utility of running a RAT curriculum entirely in a virtual format. Topics: Residents-as-teachers, distance learning, role-playing, virtual curriculum, video conferencing, One Minute Preceptor, feedback, "What if?" game, Aunt Minnie, SPIT, activated demonstration, self-directed teaching tools, teaching scripts, Post-It Pearls.

A redundant hydraulic function of root hairs in barley plants grown in hydroponics.

The root hair-less brb of Hordeum vulgare L. (bald root barley) mutant was used to assess the significance that root hairs have for the hydraulic properties of roots and response to a limited supply of mineral nutrients in plants grown on hydroponics. The barley brb mutant and its parent wild-type (H. vulgare cv. Pallas) were grown under nutrient sufficient control conditions, and under conditions of low supply of P and N. Plants were analysed when they were 14-18 days old. Root hydraulic conductivity (Lp) was determined for excised root systems and intact transpiring plants, and cell Lp was determined through cell pressure probe measurements. The formation of Casparian bands and suberin lamellae was followed through staining of cross-sections. The presence or absence of root hairs had no effect on the overall hydraulic response of plants to nutritional treatments. Root and cell Lp did not differ between the two genotypes. The most apparent difference between brb and wild-type plants was the consistently reduced formation of apoplastic barriers in brb plants. Any hydraulic function of root hairs can be redundant in barley, at least under the hydroponic conditions tested.

Recombinant Newcastle Disease Virus Immunotherapy Drives Oncolytic Effects and Durable Systemic Antitumor Immunity.

A recombinant Newcastle Disease Virus (NDV), encoding either a human (NDVhuGM-CSF, MEDI5395) or murine (NDVmuGM-CSF) GM-CSF transgene, combined broad oncolytic activity with the ability to significantly modulate genes related to immune functionality in human tumor cells. Replication in murine tumor lines was significantly diminished relative to human tumor cells. Nonetheless, intratumoral injection of NDVmuGM-CSF conferred antitumor effects in three syngeneic models in vivo; with efficacy further augmented by concomitant treatment with anti-PD-1/PD-L1 or T-cell agonists. Ex vivo immune profiling, including T-cell receptor sequencing, revealed profound immune-contexture changes consistent with priming and potentiation of adaptive immunity and tumor microenvironment (TME) reprogramming toward an immune-permissive state. CRISPR modifications rendered CT26 tumors significantly more permissive to NDV replication, and in this setting, NDVmuGM-CSF confers immune-mediated effects in the noninjected tumor in vivo Taken together, the data support the thesis that MEDI5395 primes and augments cell-mediated antitumor immunity and has significant utility as a combination partner with other immunomodulatory cancer treatments.

UME-to-GME PandEMonium in COVID-19: Large-Scale Implementation of a Virtual ACGME Milestone-Based Curriculum for Senior Medical Students Matched Into Emergency Medicine.

BACKGROUND: The COVID-19 pandemic displaced newly matched emergency medicine "pre-interns" from in-person educational experiences at the end of medical school. This called for novel remote teaching modalities. OBJECTIVE: This study assesses effectiveness of a multisite Accreditation Council for Graduate Medical Education (ACGME) sub-competency-based curricular implementation on Slack during the first wave of the COVID-19 pandemic in the United States. METHODS: Emergency medicine residency programs were recruited via national organization listservs. Programs designated instructors to manage communications and teaching for the senior medical students who had matched to their programs (pre-interns) in spring/summer 2020. Pre- and post-surveys of trainees and instructors assessed perceived preparedness for residency, perceived effectiveness of common virtual educational modalities, and concern for the pandemic's effects on medical education utilizing a Likert scale of 1 (very unconcerned) to 5 (very concerned). Data were analyzed using descriptive statistics and the t test. RESULTS: Of 276 possible residency programs, 28 enrolled. Of 324 possible pre-interns, 297 (91.7%) completed pre-surveys in April/May and 249 (76.9%) completed post-surveys in June/July. The median weeks since performing a physical examination was 8 (IQR 7-12), since attending in-person didactics was 10 (IQR 8-15) and of rotation displacement was 4 (IQR 2-6). Perceived preparedness increased both overall and for 14 of 21 ACGME Milestone topics taught. Instructors reported higher mean concern (4.32, 95% CI 4.23-4.41) than pre-interns (2.88, 95% CI 2.74-3.02) regarding the pandemic's negative effects on medical education. CONCLUSIONS: Pre-interns reported improvements in residency preparedness after participating in this ACGME sub-competency-based curriculum on Slack.

Unique receptor repertoire in mouse uterine NK cells.

Uterine NK (uNK) cells are a prominent feature of the uterine mucosa and regulate placentation. NK cell activity is regulated by a balance of activating and inhibitory receptors, however the receptor repertoire of mouse uNK cells is unknown. We describe herein two distinct subsets of CD3(-)CD122(+) NK cells in the mouse uterus (comprising decidua and mesometrial lymphoid aggregate of pregnancy) at mid-gestation: a small subset indistinguishable from peripheral NK cells, and a larger subset that expresses NKp46 and Ly49 receptors, but not NK1.1 or DX5. This larger subset reacts with Dolichus biflores agglutinin, a marker of uNK cells in the mouse, and is adjacent to the invading trophoblast. By multiparametric analysis we show that the phenotype of uNK cells is unique and unprecedented in terms of adhesion, activation, and MHC binding potential. Thus, the Ly49 repertoire and the expression of other differentiation markers strikingly distinguish uNK cells from peripheral NK cells, suggesting that a selection process shapes the receptor repertoire of mouse uNK cells.

Intratumoral IL12 mRNA Therapy Promotes TH1 Transformation of the Tumor Microenvironment.

PURPOSE: While immune checkpoint inhibitors such as anti-PD-L1 are rapidly becoming the standard of care in the treatment of many cancers, only a subset of treated patients have long-term responses. IL12 promotes antitumor immunity in mouse models; however, systemic recombinant IL12 had significant toxicity and limited efficacy in early clinical trials. EXPERIMENTAL DESIGN: We therefore designed a novel intratumoral IL12 mRNA therapy to promote local IL12 tumor production while mitigating systemic effects. RESULTS: A single intratumoral dose of mouse (m)IL12 mRNA induced IFNγ and CD8+ T-cell-dependent tumor regression in multiple syngeneic mouse models, and animals with a complete response demonstrated immunity to rechallenge. Antitumor activity of mIL12 mRNA did not require NK and NKT cells. mIL12 mRNA antitumor activity correlated with TH1 tumor microenvironment (TME) transformation. In a PD-L1 blockade monotherapy-resistant model, antitumor immunity induced by mIL12 mRNA was enhanced by anti-PD-L1. mIL12 mRNA also drove regression of uninjected distal lesions, and anti-PD-L1 potentiated this response. Importantly, intratumoral delivery of mRNA encoding membrane-tethered mIL12 also drove rejection of uninjected lesions with very limited circulating IL12p70, supporting the hypothesis that local IL12 could induce a systemic antitumor immune response against distal lesions. Furthermore, in ex vivo patient tumor slice cultures, human IL12 mRNA (MEDI1191) induced dose-dependent IL12 production, downstream IFNγ expression and TH1 gene expression. CONCLUSIONS: These data demonstrate the potential for intratumorally delivered IL12 mRNA to promote TH1 TME transformation and robust antitumor immunity.See related commentary by Cirella et al., p. 6080.