Augmenting TCR signal strength and ICOS costimulation results in metabolically fit and therapeutically potent human CAR Th17 cells.

IL-17-producing antigen-specific human T cells elicit potent antitumor activity in mice. Yet, refinement of this approach is needed to position it for clinical use. While activation signal strength regulates IL-17 production by CD4+ T cells, the degree to which T cell antigen receptor (TCR) and costimulation signal strength influences Th17 immunity remains unknown. We discovered that decreasing TCR/costimulation signal strength by incremental reduction of αCD3/costimulation beads progressively altered Th17 phenotype. Moreover, Th17 cells stimulated with αCD3/inducible costimulator (ICOS) beads produced more IL-17A, IFNγ, IL-2, and IL-22 than those stimulated with αCD3/CD28 beads. Compared with Th17 cells stimulated with the standard, strong signal strength (three beads per T cell), Th17 cells propagated with 30-fold fewer αCD3/ICOS beads were less reliant on glucose and favored the central carbon pathway for bioenergetics, marked by abundant intracellular phosphoenolpyruvate (PEP). Importantly, Th17 cells stimulated with weak αCD3/ICOS beads and redirected with a chimeric antigen receptor that recognizes mesothelin were more effective at clearing human mesothelioma. Less effective CAR Th17 cells generated with high αCD3/ICOS beads were rescued by overexpressing phosphoenolpyruvate carboxykinase 1 (PCK1), a PEP regulator. Thus, Th17 therapy can be improved by using fewer activation beads during manufacturing, a finding that is cost effective and directly translatable to patients.

Developing Nurses and Interdisciplinary Teams in Caring Science.

This program evaluation assessed a caring science program's impact on nurse and interdisciplinary professionals' self-reported caring, compassion satisfaction, and intent to leave at an academic-affiliated community hospital. A 3-session program resulted in self-caring and intent to leave significant increases at 60 days post intervention. Findings demonstrated caring science interventions alone are insufficient to impact staff engagement and intent to leave. Further actions for organizational culture changes are discussed.

Perfusion Imaging for Endovascular Thrombectomy in Acute Ischemic Stroke Is Associated With Improved Functional Outcomes in the Early and Late Time Windows.

BACKGROUND: The impact on clinical outcomes of patient selection using perfusion imaging for endovascular thrombectomy (EVT) in patients with acute ischemic stroke presenting beyond 6 hours from onset remains undetermined in routine clinical practice. METHODS: Patients from a national stroke registry that underwent EVT selected with or without perfusion imaging (noncontrast computed tomography/computed tomography angiography) in the early (<6 hours) and late (6-24 hours) time windows, between October 2015 and March 2020, were compared. The primary outcome was the ordinal shift in the modified Rankin Scale score at hospital discharge. Other outcomes included functional independence (modified Rankin Scale score ≤2) and in-hospital mortality, symptomatic intracerebral hemorrhage, successful reperfusion (Thrombolysis in Cerebral Infarction score 2b-3), early neurological deterioration, futile recanalization (modified Rankin Scale score 4-6 despite successful reperfusion) and procedural time metrics. Multivariable analyses were performed, adjusted for age, sex, baseline stroke severity, prestroke disability, intravenous thrombolysis, mode of anesthesia (Model 1) and including EVT technique, balloon guide catheter, and center (Model 2). RESULTS: We included 4249 patients, 3203 in the early window (593 with perfusion versus 2610 without perfusion) and 1046 in the late window (378 with perfusion versus 668 without perfusion). Within the late window, patients with perfusion imaging had a shift towards better functional outcome at discharge compared with those without perfusion imaging (adjusted common odds ratio [OR], 1.45 [95% CI, 1.16-1.83]; P=0.001). There was no significant difference in functional independence (29.3% with perfusion versus 24.8% without; P=0.210) or in the safety outcome measures of symptomatic intracerebral hemorrhage (P=0.53) and in-hospital mortality (10.6% with perfusion versus 14.3% without; P=0.053). In the early time window, patients with perfusion imaging had significantly improved odds of functional outcome (adjusted common OR, 1.51 [95% CI, 1.28-1.78]; P=0.0001) and functional independence (41.6% versus 33.6%, adjusted OR, 1.31 [95% CI, 1.08-1.59]; P=0.006). Perfusion imaging was associated with lower odds of futile recanalization in both time windows (late: adjusted OR, 0.70 [95% CI, 0.50-0.97]; P=0.034; early: adjusted OR, 0.80 [95% CI, 0.65-0.99]; P=0.047). CONCLUSIONS: In this real-world study, acquisition of perfusion imaging for EVT was associated with improvement in functional disability in the early and late time windows compared with nonperfusion neuroimaging. These indirect comparisons should be interpreted with caution while awaiting confirmatory data from prospective randomized trials.

Ex vivo blockade of PI3K gamma or delta signaling enhances the antitumor potency of adoptively transferred CD8+ T cells.

Adoptive T cell transfer therapy induces objective responses in patients with advanced malignancies. Despite these results, some individuals do not respond due to the generation of terminally differentiated T cells during the expansion protocol. As the gamma and delta catalytic subunits in the PI3K pathway are abundant in leukocytes and involved in cell activation, we posited that blocking both subunits ex vivo with the inhibitor IPI-145 would prevent their differentiation, thereby increasing antitumor activity in vivo. However, IPI-145 treatment generated a product with reduced antitumor activity. Instead, T cells inhibited of PI3Kγ (IPI-549) or PI3Kδ (CAL-101 or TGR-1202) alone were more potent in vivo. While T cells coinhibited of PI3Kγ and PI3Kδ were less differentiated, they were functionally impaired, indicated by reduced production of effector cytokines after antigenic re-encounter and decreased persistence in vivo. Human CAR T cells expanded with either a PI3Kγ or PI3Kδ inhibitor possessed a central memory phenotype compared to vehicle cohorts. We also found that PI3Kδ-inhibited CARs lysed human tumors in vitro more effectively than PI3Kγ-expanded or traditionally expanded CAR T cells. Our data imply that sole blockade of PI3Kγ or PI3Kδ generates T cells with remarkable antitumor properties, a discovery that has substantial clinical implications.

Same day embolisation followed by microsurgical resection of brain arteriovenous malformations: a single centre early experience.

INTRODUCTION: The purpose of this paper is to report our experience of treating cerebral arteriovenous malformations (AVM) in adults with pre-operative embolisation and microsurgical resection on the same day during a single anaesthetic at a single centre between April 2016 and December 2018. We included both elective AVM and AVM that had bled acutely. METHODS: We retrospectively analysed data from patients with cerebral AVMs who underwent embolisation followed by microsurgical resection on the same day at a single neurosurgical centre. PRIMARY ENDPOINTS INCLUDED: Total procedure time (embolisation and microsurgical resection), procedure finish time, intra-operative blood loss, degree of nidus obliteration on postoperative angiography, intensive care unit (ICU) stay, total stay at the neurosurgical centre and modified Rankin Score (pre- and post-procedure). RESULTS: •Nineteen patients underwent same-day pre-operative embolisation and microsurgical resection over the 32-month period. The average patient age was 40 years (range 19-66 years). One patient had undergone a prior attempt at embolisation and one patient previously had sterotactic radiosurgery (STRS). •Thirteen of the AVM were in the dominant hemisphere and six in the non-dominant hemisphere. Sixteen AVM were located supratentorially and three were in the posterior fossa. Spetzler-Martin grades included 4 grade 1, 10 grade 2, 4 grade 3 and 1 grade 4. •The average blood loss intra-operatively was 289 mls. •The average list finish time was 19:56 (range 15:10-00:00). •Seventeen patients had 100% nidus obliteration on post-operative digital subtraction angiography, one patient had a small remnant and was referred to STRS and one patient died in the ICU post operatively. CONCLUSION: Overall, the authors believe same-day embolisation and microsurgical resection represents a safe treatment strategy. The technique minimises hemorrhagic complications from delayed venous occlusion and avoids multiple anaesthetics and hospital admissions.

Progressively Enhancing Stemness of Adoptively Transferred T Cells with PI3Kδ Blockade Improves Metabolism and Antitumor Immunity.

Generating stem-like memory T cells (TSCM) is a potential strategy to improve adoptive immunotherapy. Elucidating optimal ways to modulate signaling pathways that enrich TSCM properties could identify approaches to achieve this goal. We discovered herein that blocking the PI3Kδ pathway pharmaceutically to varying degrees can generate T cells with increasingly heightened stemness properties, based on the progressive enrichment of the transcription factors Tcf1 and Lef1. T cells with enhanced stemness features exhibited metabolic plasticity, marked by improved mitochondrial function and glucose uptake after tumor recognition. Conversely, T cells with low or medium stemness were less metabolically dynamic, vulnerable to antigen-induced cell death, and expressed more inhibitory checkpoint receptors. Only T-cell receptor-specific or chimeric antigen receptor (CAR)-specific T cells with high stemness persisted in vivo and mounted protective immunity to tumors. Likewise, the strongest level of PI3Kδ blockade in vitro generated human tumor-infiltrating lymphocytes and CAR T cells with elevated stemness properties, in turn bolstering their capacity to regress human solid tumors. The stemness level of T cells in vitro was important, ultimately impacting their efficacy in mice bearing three distinct solid tumors. Lef1 and Tcf1 sustained antitumor protection by donor high CD8+ TSCM or CD4+ Th17SCM, as deletion of either one compromised the therapeutic efficacy. Collectively, these findings highlight the importance of strategic modulation of PI3Kδ signaling in T cells to induce stemness and lasting protective responses to solid tumors. SIGNIFICANCE: Elevating T-cell stemness by progressively blocking PI3Kδ signaling during ex vivo manufacturing of adoptive cell therapies alters metabolic and functional properties to enhance antitumor immunity dependent on Tcf1 and Lef1.

Motivation, management, and mastery: a theory of resilience in the context of HIV infection.

BACKGROUND: Clients with HIV infection have been conceptualized as a resilient population. Although a few researchers have documented resilience among clients with HIV infection, a theory of resilience in the context of HIV infection has not been developed. The purpose of this study was to describe the process by which resilience occurs for clients in the context of HIV infection. METHOD: Grounded theory methodology was used to sample and analyze data from 15 qualitative interviews with adults with HIV infection. Data were collected until saturation was reached. RESULTS: A theory, motivation, management, and mastery, a description of the process by which resilience occurs in the context of HIV infection, emerged from the data. CONCLUSION: Many clients living with HIV infection are resilient, despite the physical, psychological, and social challenges of this chronic illness. Nursing interventions to promote resilience among clients with HIV infection should be directed toward identification of client motivation factors and disease management strategies that may influence health outcomes of people living with HIV infection.

Voices of Musicians: Virtual Live Bedside Music Concerts in Inpatient Care.

The COVID-19 pandemic presented unprecedented challenges to patients, family members, and healthcare staff that resulted in increased stress and isolation and decreased quality of life. We evaluate the impact of a novel virtual concert program, the Vital Sounds Initiative (VSI) of Project: Music Heals Us (PMHU), which began at the beginning of the pandemic to combat patient isolation and provide employment to professional musicians. Using a qualitative analysis of VSI data, we examined post-concert written responses by musicians. These responses were coded by independent coders via inductive coding and thematic analysis. Between 7 April 2020 and 20 July 2022, 192 musicians played 2203 h of music for 11,222 audience members in 39 care facilities nationwide. A total of 114 musicians submitted a total of 658 responses. Three main themes (with corresponding subthemes) arose: (1) Patient Experience; (2) Musician Experience; (3) Caregiver (family or staff) Experience. The responses offered valuable insight into the overwhelmingly positive aspects of the virtual concerts. Overall, we found that VSI favorably impacts individuals at every level, including the patients, musician, and caregivers. These findings provide preliminary evidence for the benefits of virtual music concerts. Upscaling similar virtual music interventions/programs should be considered.

The degree of T cell stemness differentially impacts the potency of adoptive cancer immunotherapy in a Lef-1 and Tcf-1 dependent manner.

Generating stem memory T cells (T SCM ) is a key goal for improving cancer immunotherapy. Yet, the optimal way to modulate signaling pathways that enrich T SCM properties remains elusive. Here, we discovered that the degree to which the PI3Kδ pathway is blocked pharmaceutically can generate T cells with differential levels of stemness properties. This observation was based on the progressive enrichment of transcriptional factors of stemness (Tcf-1 and Lef-1). Additional investigation revealed that T cells with high stemness features had enhanced metabolic plasticity, marked by heightened mitochondrial function and glucose uptake. Conversely, T cells with low or medium features of stemness expressed more inhibitory checkpoint receptors (Tim-3, CD39) and were vulnerable to antigen-induced cell death. Only TCR-antigen specific T cells with high stemness persisted following adoptive transfer in vivo and mounted protective immunity to melanoma tumors. Likewise, the strongest level of PI3Kδ blockade in vitro generated human tumor infiltrating lymphocytes (TILs) and CAR T cells with heightened stemness properties, in turn bolstering their capacity to regress human mesothelioma tumors. We find that the level of stemness T cells possess in vitro differentially impacts their potency upon transfer in three tumor models. Mechanistically, both Lef-1 and Tcf-1 sustain anti-tumor protection by high T SCM , as deletion of either one compromised cellular therapy. Collectively, these findings highlight the therapeutic potential of carefully modulating PI3Kδ signaling in T cells to confer high stemness and mediate protective responses to solid tumors.

Adhesion analysis via a tumor vasculature-like microfluidic device identifies CD8+ T cells with enhanced tumor homing to improve cell therapy.

CD8+ T cell recruitment to the tumor microenvironment is critical for the success of adoptive cell therapy (ACT). Unfortunately, only a small fraction of transferred cells home to solid tumors. Adhesive ligand-receptor interactions have been implicated in CD8+ T cell homing; however, there is a lack of understanding of how CD8+ T cells interact with tumor vasculature-expressed adhesive ligands under the influence of hemodynamic flow. Here, the capacity of CD8+ T cells to home to melanomas is modeled ex vivo using an engineered microfluidic device that recapitulates the hemodynamic microenvironment of the tumor vasculature. Adoptively transferred CD8+ T cells with enhanced adhesion in flow in vitro and tumor homing in vivo improve tumor control by ACT in combination with immune checkpoint blockade. These results show that engineered microfluidic devices can model the microenvironment of the tumor vasculature to identify subsets of T cells with enhanced tumor infiltrating capabilities, a key limitation in ACT.

Implantation of Large Diameter (5.5-6 mm) Derivo Embolization Devices for the Treatment of Cerebral Aneurysms.

BACKGROUND: The efficacy of flow diverters is dependent upon robust wall apposition in the parent artery. Usage in large caliber cerebral vessels has therefore been limited as few implants with diameters > 5 mm exist. We present our initial experience in treating cerebral aneurysms using the 5.5 mm and 6 mm diameter implants of the Derivo embolization device (DED). METHODS: Our prospectively maintained institutional database was reviewed to identify patients in whom a > 5 mm DED was implanted between November 2016 and February 2021. The primary efficacy outcome was complete or near-complete aneurysm occlusion at 6 months (O'Kelly-Marotta, OKM, C-D, adapted for magnetic resonance angiography). Safety outcomes included 30-day major morbidity defined as modified Rankin Score (mRS) 3-5, mortality, serious adverse events and procedural complications. RESULTS: A total of 21 large diameter DEDs were deployed in 18 patients (age 59.5 ± 14.1 years), harboring 19 unruptured aneurysms. Of the aneurysms 14 (73.7%) were saccular in morphology (sac diameter 10.9 ± 5.5 mm, neck diameter 6.8 ± 3.1 mm), 3 (15.8%) aneurysms were dissecting, 1 (5.3%) iatrogenic pseudoaneurysm and 1 (5.3%) fusiform. Aneurysm locations were: ICA (internal carotid artery) (n = 17); (7 cavernous, 4 paraophthalmic, 2 paraclinoid, 1 petrous, 2 communicating, 1 cervical); vertebrobasilar (n = 2). Adjunct stenting to optimize proximal wall apposition was undertaken in 5 (27.8%) patients. At 6 months 75% of patients followed-up met the primary efficacy endpoint (OKM C-D). There were no serious adverse events, 30-day major morbidity (mRS 3-5) or mortality. CONCLUSION: Implantation of large diameter (5.5 mm and 6 mm) DEDs into capacious cerebral vessels to treat a range of complex aneurysms is safe and technically feasible but may require adjunct stenting to optimize proximal wall apposition. Short-term efficacy of this device subset is comparable to previous DED and other flow diverter studies. Long-term follow-up and comparative studies are required for further assessment.

Prognostic factors following resection of intracranial metastases.

Background: The aim of this study was to identify prognostic factors associated with resection of intracranial metastases. Methods: A retrospective case series including patients who underwent resection of cranial metastases from March 2014 to April 2021 at a single center. This identified 112 patients who underwent 124 resections. The median age was 65 years old (24-84) and the most frequent primary cancers were nonsmall cell lung cancer (56%), breast adenocarcinoma (13%), melanoma (6%), and colorectal adenocarcinoma (6%). Postoperative MRI with contrast was performed within 48 hours in 56% of patients and radiation treatment was administered in 41%. GraphPad Prism 9.2.0 was used for the survival analysis. Results: At the time of data collection, 23% were still alive with a median follow-up of 1070 days (68-2484). The 30- and 90-day, and 1- and 5-year overall survival rates were 93%, 83%, 35%, and 17%, respectively. The most common causes of death within 90 days were as follows: unknown (32%), systemic or intracranial disease progression (26%), and pneumonia (21%). Age and extent of neurosurgical resection were associated with overall survival (P < 0.05). Patients aged >70 had a median survival of 5.4 months compared with 9.7, 11.4, and 11.4 for patients <50, 50-59, and 60-69, respectively. Gross-total resection achieved an overall survival of 11.8 months whereas sub-total, debulking, and unclear extent of resection led to a median survival of 5.7, 7.0, and 9.0 months, respectively. Conclusion: Age and extent of resection are potential predictors of long-term survival.

B cells imprint adoptively transferred CD8+ T cells with enhanced tumor immunity.

BACKGROUND: Adoptive T cell transfer (ACT) therapy improves outcomes in patients with advanced malignancies, yet many individuals relapse due to the infusion of T cells with poor function or persistence. Toll-like receptor (TLR) agonists can invigorate antitumor T cell responses when administered directly to patients, but these responses often coincide with toxicities. We posited that TLR agonists could be repurposed ex vivo to condition T cells with remarkable potency in vivo, circumventing TLR-related toxicity. METHODS: In this study we investigated how tumor-specific murine CD8+ T cells and human tumor infiltrating lymphocytes (TILs) are impacted when expanded ex vivo with the TLR9 agonist CpG. RESULTS: Herein we reveal a new way to reverse the tolerant state of adoptively transferred CD8+ T cells against tumors using TLR-activated B cells. We repurposed the TLR9 agonist, CpG, commonly used in the clinic, to bolster T cell-B cell interactions during expansion for ACT. T cells expanded ex vivo from a CpG-treated culture demonstrated potent antitumor efficacy and prolonged persistence in vivo. This antitumor efficacy was accomplished without in vivo administration of TLR agonists or other adjuvants of high-dose interleukin (IL)-2 or vaccination, which are classically required for effective ACT therapy. CpG-conditioned CD8+ T cells acquired a unique proteomic signature hallmarked by an IL-2RαhighICOShighCD39low phenotype and an altered metabolic profile, all reliant on B cells transiently present in the culture. Likewise, human TILs benefitted from expansion with CpG ex vivo, as they also possessed the IL-2RαhighICOShighCD39low phenotype. CpG fostered the expansion of potent CD8+ T cells with the signature phenotype and antitumor ability via empowering a direct B-T cell interaction. Isolated B cells also imparted T cells with the CpG-associated phenotype and improved tumor immunity without the aid of additional antigen-presenting cells or other immune cells in the culture. CONCLUSIONS: Our results demonstrate a novel way to use TLR agonists to improve immunotherapy and reveal a vital role for B cells in the generation of potent CD8+ T cell-based therapies. Our findings have immediate implications in the clinical treatment of advanced solid tumors.

Substance abuse, violence, HIV, and depression: an underlying syndemic factor among Latinas.

BACKGROUND: Evidence from the literature suggests that substance abuse, violence, HIV risk, depressive symptoms, and underlying socioeconomic conditions are tied intrinsically to health disparities among Latinas. Although these health and social conditions appear to comprise a syndemic, an underlying phenomenon disproportionately accounting for the burden of disease among marginalized groups, these hypothesized relationships have not been formally tested. OBJECTIVES: The aim of this study was to assess (a) if substance abuse, violence, HIV risk, and depressive symptoms comprised a syndemic and (b) if this syndemic was related to socioeconomic disadvantage among Latinas. METHODS: Baseline assessment data from a randomized controlled community trial testing the efficacy of an HIV risk reduction program for adult Latinas (n = 548) were used to measure demographic variables, substance abuse, violence, risk for HIV, and depressive symptoms. Structural equation modeling was used to test a single underlying syndemic factor model and any relation to socioeconomic disadvantage. RESULTS: The results of this study support the idea that HIV risk, substance abuse, violence, and depressive symptoms comprise a syndemic, χ(27) = 53.26, p < .01 (relative χ = 1.97, comparative fit index = .91, root mean square error of approximation = .04). In addition, in limited accord with theory, this factor was related to 2 measures of socioeconomic disadvantage, percentage of years in the United States (b = 7.55, SE = 1.53, p < .001) and education (b = -1.98, SE = .87, p < .05). DISCUSSION: The results of this study could be used to guide public health programs and policies targeting behavioral health disparity conditions among Latinos and other vulnerable populations. Further study of the influence of gender-role expectations and community-level socioeconomic indicators may provide additional insight into this syndemic.

A syndemic model of substance abuse, intimate partner violence, HIV infection, and mental health among Hispanics.

Hispanics are disproportionately affected by substance abuse, HIV infection, intimate partner violence, and mental health conditions. To address health disparities among Hispanics and other vulnerable groups, it is necessary to understand the complex interactions between health conditions clustering together (e.g., substance abuse, intimate partner violence, and HIV) and the social ecology in which these conditions exist. A syndemic orientation, a consideration of clustering epidemics and common individual, relationship, cultural, and socioenvironmental factors linking these conditions, may be helpful in developing comprehensive models that expand our ability to understand and address health disparities. The purpose of this paper is to introduce a Syndemic Model of Substance Abuse, Intimate Partner Violence, HIV Infection, and Mental Health among Hispanics, and provide evidence from the research literature to support the central relationships and risk and protective factors (i.e., potential links between conditions) depicted by the model. The development and evaluation of interventions aimed at the prevention of substance abuse, intimate partner violence, HIV/AIDS, and mental health problems as a syndemic affecting Hispanics is urgently needed. Public health nurses can initiate this endeavor with the guidance of a Syndemic Model.

Fundamentals of T Cell Metabolism and Strategies to Enhance Cancer Immunotherapy.

Emerging reports show that metabolic pathways can be targeted to enhance T cell-mediated immunity to tumors. Yet, tumors consume key metabolites in the host to survive, thus robbing T cells of these nutrients to function and thrive. T cells are often deprived of basic building blocks for energy in the tumor, including glucose and amino acids needed to proliferate or produce cytotoxic molecules against tumors. Immunosuppressive molecules in the host further compromise the lytic capacity of T cells. Moreover, checkpoint receptors inhibit T cell responses by impairing their bioenergetic potential within tumors. In this review, we discuss the fundamental metabolic pathways involved in T cell activation, differentiation and response against tumors. We then address ways to target metabolic pathways to improve the next generation of immunotherapies for cancer patients.

Modeling ex vivo tumor-infiltrating lymphocyte expansion from established solid malignancies.

Adoptive transfer of tumor-infiltrating lymphocytes (TIL) elicits the regression of metastatic malignancies, yet a low proportion of patients achieve complete durable responses. The high incidence of relapse in these patients highlights the need to better understand mechanisms of tumor escape from T cell control. While melanoma has provided the foundation for developing TIL therapy, much less is known about TIL efficacy and relapse in other malignancies. We sought to investigate TIL characteristics in mouse tumors which have not been studied in this setting. Here, we expanded murine TIL ex vivo in IL-2 from fragments of multiple tumor models, including oral cavity cancer models of varying immunogenicity. Additionally, TIL was expanded from pmel-1 mice bearing B16F10 melanoma, yielding an enriched population of tumor-infiltrating TCR transgenic T cells. Murine TIL are similar to human TIL in that they express high levels of inhibitory receptors (PD-1, Tim-3, etc.) and can be expanded ex vivo in IL-2 extensively. Of clinical relevance, we draw parallels between murine and human oral cavity cancer TIL, evaluating relationships between inhibitory receptor expression and function. This platform can be used by labs even in the absence of clinical specimens or clean cell facilities and will be important to more broadly understand TIL phenotypes across many different malignancies.

NEAT1 is essential for metabolic changes that promote breast cancer growth and metastasis.

Accelerated glycolysis is the main metabolic change observed in cancer, but the underlying molecular mechanisms and their role in cancer progression remain poorly understood. Here, we show that the deletion of the long noncoding RNA (lncRNA) Neat1 in MMTV-PyVT mice profoundly impairs tumor initiation, growth, and metastasis, specifically switching off the penultimate step of glycolysis. Mechanistically, NEAT1 directly binds and forms a scaffold bridge for the assembly of PGK1/PGAM1/ENO1 complexes and thereby promotes substrate channeling for high and efficient glycolysis. Notably, NEAT1 is upregulated in cancer patients and correlates with high levels of these complexes, and genetic and pharmacological blockade of penultimate glycolysis ablates NEAT1-dependent tumorigenesis. Finally, we demonstrate that Pinin mediates glucose-stimulated nuclear export of NEAT1, through which it exerts isoform-specific and paraspeckle-independent functions. These findings establish a direct role for NEAT1 in regulating tumor metabolism, provide new insights into the Warburg effect, and identify potential targets for therapy.

Redox-active ligand-mediated oxidative addition and reductive elimination at square planar cobalt(III): multielectron reactions for cross-coupling.

Square planar cobalt(III) complexes with redox-active amidophenolate ligands are strong nucleophiles that react with alkyl halides, including CH(2)Cl(2), under gentle conditions to generate stable square pyramidal alkylcobalt(III) complexes. The net electrophilic addition reactions formally require 2e(-) oxidation of the metal fragment, but there is no change in metal oxidation state because the reaction proceeds with 1e(-) oxidation of each amidophenolate ligand. Although the four-coordinate complexes are very strong nucleophiles, they are mild outer-sphere reductants. Accordingly, addition of alkyl- or phenylzinc halides to the five-coordinate organometallic complexes regenerates the square planar starting materials and extrudes C-C coupling products. The net 2e(-) reductive elimination reaction also occurs without a oxidation state change at the cobalt(III) center. Together these reactions comprise a complete, well-defined cycle for cobalt Negishi-like cross-coupling of alkyl halides with organozinc reagents.