Dual-regulated lentiviral vector for gene therapy of X-linked chronic granulomatosis

Regulated transgene expression may improve the safety and efficacy of hematopoietic stem cell (HSC) gene therapy. Clinical trials for X-linked chronic granulomatous disease (X-CGD) employing gammaretroviral vectors were limited by insertional oncogenesis or lack of persistent engraftment. Our novel strategy, based on regulated lentiviral vectors (LV), targets gp91(phox) expression to the differentiated myeloid compartment while sparing HSC, to reduce the risk of genotoxicity and potential perturbation of reactive oxygen species levels. Targeting was obtained by a myeloid-specific promoter (MSP) and posttranscriptional, microRNA-mediated regulation. We optimized both components in human bone marrow (BM) HSC and their differentiated progeny in vitro and in a xenotransplantation model, and generated therapeutic gp91(phox) expressing LVs for CGD gene therapy. All vectors restored gp91(phox) expression and function in human X-CGD myeloid cell lines, primary monocytes, and differentiated myeloid cells. While unregulated LVs ectopically expressed gp91(phox) in CD34(+) cells, transcriptionally and posttranscriptionally regulated LVs substantially reduced this off-target expression. X-CGD mice transplanted with transduced HSC restored gp91(phox) expression, and MSP-driven vectors maintained regulation during BM development. Combining transcriptional (SP146.gp91-driven) and posttranscriptional (miR-126-restricted) targeting, we achieved high levels of myeloid-specific transgene expression, entirely sparing the CD34(+) HSC compartment. This dual-targeted LV construct represents a promising candidate for further clinical development

The prospects for the SARS‐CoV‐2 pandemic in Africa

On December 31, 2019, the Chinese government officially announced the identification of a new type of coronavirus (SARS‐CoV‐2) as the etiological cause of a severe acute respiratory syndrome in Wuhan city, Hubei Province. Over the next weeks, SARS‐CoV‐2 caused a global pandemic as officially declared by the WHO on March 11, 2020, with confirmed cases and deaths in more than 166 countries. We are experiencing a worldwide phenomenon of unprecedented social and economic consequences. Since the beginning of the COVID‐19 outbreak, there have been fears that the epidemic could strongly impact weaker healthcare systems in poor‐resource settings, especially in Sub‐Saharan Africa (SSA). The 2 million Chinese nationals that live and work in Africa could potentially contribute to the spread of COVID‐19 on the continent

USE OFTHE NEW INTEGRATED TABLE MOTION FOR THE DA VINCI XI SURGICAL SYSTEM IN ABDOMINAL SURGICAL PROCEDURES

PURPOSE Integrated Table Motion (ITM) for the da Vinci Xi Surgical System is a new feature comprising of a Trumpf TS7000dV Operating Table, that communicates wirelessly with the da Vinci Xi. The feature allows the surgical staff to reposition the patient without undocking the robot from the patient and without removing instruments from inside the abdomen. ITM has been specifically developed to improve multiquadrant robotic surgery. We herein present the first human use of this device across multiple surgical disciplines in the EU. METHODS Between May and October 2015 one of the first human use of ITM was conducted in a post market study in the EU in which 30 cases across different specialties were prospectively enrolled. We included general surgery procedures, urological procedures and gynecological procedures. Variables examined included patient characteristics and intraoperative data. Primary end-points were: ITM efficacy, safety and efficiency. For these reasons we evaluated the number of table moves per case, duration of each table move, table positions attained , reasons for moving the table and the states of instruments and endoscope during table move (inserted or removed). We also evaluated the safety of ITM by recording occurrence of adverse events related to the use of ITM. RESULTS Twelve patients underwent general surgery procedures (six cases of anterior rectal resection (ARR) with TME, 3 cases of right hemicolectomy, a case of subtotal gastrectomy, a case of hepatic resection and a case of ventral hernia repair. Gynecological procedures included five cases of hysterectomy, a case of repair of rectal prolapse and a case of uterine prolapse. Urological procedures included seven cases of prostatectomy, a case of nephrectomy, two cases of partial nephrectomy and a case of pyeloplasty. The mean ITM moves during the colorectal procedures was 3, while ITM was moved two times for repair of rectal prolapsed, three times during subtotal gastrectomy and only one time during liver resection and ventral hernia repair, resulting in 35 instances of table moves in 13 procedures. The mean ITM moves during the repair of uterine prolapse was 5, while ITM was moved on average 3.4 times during hysterectomy, resulting in 22 instances of table moves in 6 procedures. The mean ITM moves during prostatectomy was 3.1, while ITM was moved 2 times during partial and total nephrectomy and pyeloplasty, resulting in 30 instances of table moves in 11 procedures. Majority of moves (>70%) took less than 2 minutes to complete. The primary reason for using ITM was to gain internal exposure in 82 moves (93%). The endoscope was left inserted during 91-93% of table movements, while the instruments were left inserted in 95-97% of moves. No external collisions or other problems related to the operating table were noted. There were no ITM related surgical complications or need for conversion to laparoscopy or laparotomy. There were no ITM safety-related observations and no adverse events ITM or device-related. CONCLUSIONS This preliminary study demonstrated the efficiency of ITM for the da Vinci Xi Surgical System, which enabled patient repositioning without disrupting surgical workflow by allowing the surgeon to leave instruments and the scope docked to the patient. ITM has been shown to be safe, and no adverse events related to its use were reported. Further studies can be useful to demonstrate if ITM could enable procedures or part of procedures to be done robotically that would otherwise be difficult, and if ITM could improve operative efficiency by reducing surgical operative time

Artificial intelligence-based prediction of transfusion in the intensive care unit in patients with gastrointestinal bleeding

Objective Gastrointestinal (GI) bleeding commonly requires intensive care unit (ICU) in cases of potentialhaemodynamiccompromise or likely urgent intervention. However, manypatientsadmitted to the ICU stop bleeding and do not require further intervention, including blood transfusion. The present work proposes an artificial intelligence (AI) solution for the prediction of rebleeding in patients with GI bleeding admitted to ICU. Methods A machine learning algorithm was trained and tested using two publicly available ICU databases, the Medical Information Mart for Intensive Care V.1.4 database and eICU Collaborative Research Database using freedom from transfusion as a proxy for patients who potentially did not require ICU-level care. Multiple initial observation time frames were explored using readily available data including labs, demographics and clinical parameters for a total of 20 covariates. Results The optimal model used a 5-hour observation period to achieve an area under the curve of the receiving operating curve (ROC-AUC) of greater than 0.80. The model was robust when tested against both ICU databases with a similar ROC-AUC for all. Conclusions The potential disruptive impact of AI in healthcare innovation is acknowledge, but awareness of AI-related risk on healthcare applications and current limitations should be considered before implementation and deployment. The proposed algorithm is not meant to replace but to inform clinical decision making. Prospective clinical trial validation as a triage tool is warranted.National Science Foundation (Grant NIBIB R01 EB017205

Targeted single-cell electroporation of mammalian neurons in vivo.

In order to link our knowledge of single neurons with theories of network function, it has been a long-standing goal to manipulate the activity and gene expression of identified subsets of mammalian neurons within the intact brain in vivo. This protocol describes a method for delivering plasmid DNA into single identified mammalian neurons in vivo, by combining two-photon imaging with single-cell electroporation. Surgery, mounting of a chronic recording chamber and targeted electroporation of identified neurons can be performed within 1-2 h. Stable transgene expression can reliably be induced with high success rates both in single neurons as well as in small, spatially defined networks of neurons in the cerebral cortex of rodents

IL-1β+ macrophages fuel pathogenic inflammation in pancreatic cancer

: Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with high resistance to therapies1. Inflammatory and immunomodulatory signals co-exist in the pancreatic tumour microenvironment, leading to dysregulated repair and cytotoxic responses. Tumour-associated macrophages (TAMs) have key roles in PDAC2, but their diversity has prevented therapeutic exploitation. Here we combined single-cell and spatial genomics with functional experiments to unravel macrophage functions in pancreatic cancer. We uncovered an inflammatory loop between tumour cells and interleukin-1β (IL-1β)-expressing TAMs, a subset of macrophages elicited by a local synergy between prostaglandin E2 (PGE2) and tumour necrosis factor (TNF). Physical proximity with IL-1β+ TAMs was associated with inflammatory reprogramming and acquisition of pathogenic properties by a subset of PDAC cells. This occurrence was an early event in pancreatic tumorigenesis and led to persistent transcriptional changes associated with disease progression and poor outcomes for patients. Blocking PGE2 or IL-1β activity elicited TAM reprogramming and antagonized tumour cell-intrinsic and -extrinsic inflammation, leading to PDAC control in vivo. Targeting the PGE2-IL-1β axis may enable preventive or therapeutic strategies for reprogramming of immune dynamics in pancreatic cancer