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.
We therefore designed a novel intratumoral IL12 mRNA therapy to promote local IL12 tumor production while mitigating systemic effects.
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
patient tumor slice cultures, human IL12 mRNA (MEDI1191) induced dose-dependent IL12 production, downstream IFNγ expression and TH1 gene expression.
These data demonstrate the potential for intratumorally delivered IL12 mRNA to promote TH1 TME transformation and robust antitumor immunity.
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Transcription factors play important roles in lymphopoiesis. We have previously demonstrated that Bcl11a is essential for normal lymphocyte development in the mouse embryo. We report here that, in ...the adult mouse, Bcl11a is expressed in most hematopoietic cells and is highly enriched in B cells, early T cell progenitors, common lymphoid progenitors (CLPs), and hematopoietic stem cells (HSCs). In the adult mouse, Bcl11a deletion causes apoptosis in early B cells and CLPs and completely abolishes the lymphoid development potential of HSCs to B, T, and NK cells. Myeloid development, in contrast, is not obviously affected by the loss of Bcl11a. Bcl11a regulates expression of Bcl2, Bcl2-xL, and Mdm2, which inhibits p53 activities. Overexpression of Bcl2 and Mdm2, or p53 deficiency, rescues both lethality and proliferative defects in Bcl11a-deficient early B cells and enables the mutant CLPs to differentiate to lymphocytes. Bcl11a is therefore essential for lymphopoiesis and negatively regulates p53 activities. Deletion of Bcl11a may represent a new approach for generating a mouse model that completely lacks an adaptive immune system.
Hematopoietic stem cells (HSCs) are a rare cell type with the ability of long-term self-renewal and multipotency to reconstitute all blood lineages. HSCs are typically purified from the bone marrow ...using cell surface markers. Recent studies have identified significant cellular heterogeneities in the HSC compartment with subsets of HSCs displaying lineage bias. We previously discovered that the transcription factor Bcl11a has critical functions in the lymphoid development of the HSC compartment.
In this report, we employ single-cell transcriptomic analysis to dissect the molecular heterogeneities in HSCs. We profile the transcriptomes of 180 highly purified HSCs (Bcl11a (+/+) and Bcl11a (-/-)). Detailed analysis of the RNA-seq data identifies cell cycle activity as the major source of transcriptomic variation in the HSC compartment, which allows reconstruction of HSC cell cycle progression in silico. Single-cell RNA-seq profiling of Bcl11a (-/-) HSCs reveals abnormal proliferative phenotypes. Analysis of lineage gene expression suggests that the Bcl11a (-/-) HSCs are constituted of two distinct myeloerythroid-restricted subpopulations. Remarkably, similar myeloid-restricted cells could also be detected in the wild-type HSC compartment, suggesting selective elimination of lymphoid-competent HSCs after Bcl11a deletion. These defects are experimentally validated in serial transplantation experiments where Bcl11a (-/-) HSCs are myeloerythroid-restricted and defective in self-renewal.
Our study demonstrates the power of single-cell transcriptomics in dissecting cellular process and lineage heterogeneities in stem cell compartments, and further reveals the molecular and cellular defects in the Bcl11a-deficient HSC compartment.
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.
NK cells use a variety of receptors to detect abnormal cells, including tumors and their metastases. However, in the case of melanoma, it remains to be determined what specific molecular interactions ...are involved and whether NK cells control metastatic progression and/or the route of dissemination. Here we show that human melanoma cell lines derived from LN metastases express ligands for natural cytotoxicity receptors (NCRs) and DNAX accessory molecule-1 (DNAM-1), two emerging NK cell receptors key for cancer cell recognition, but not NK group 2 member D (NKG2D). Compared with cell lines derived from metastases taken from other anatomical sites, LN metastases were more susceptible to NK cell lysis and preferentially targeted by adoptively transferred NK cells in a xenogeneic model of cell therapy. In mice, DNAM-1 and NCR ligands were also found on spontaneous melanomas and melanoma cell lines. Interference with DNAM-1 and NCRs by antibody blockade or genetic disruption reduced killing of melanoma cells. Taken together, these results show that DNAM-1 and NCRs are critical for NK cell-mediated innate immunity to melanoma cells and provide a background to design NK cell-based immunotherapeutic strategies against melanoma and possibly other tumors.
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.
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.
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.
Abstract
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.
The Residents-as-Teachers (RAT) curriculum is designed for emergency medicine (EM) residents of all years (PGY1-4).
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.
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.
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.
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.
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.
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.
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.
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.
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