Impact of continuous glucose monitoring on hospitalizations and glucose control in people with type 2 diabetes: real-world analysis.

AIM: The real-world benefits of continuous glucose monitoring (CGM) in the broad type 2 diabetes (T2D) population are not well studied. Our study evaluated the impact of CGM use on health care resource utilization over 12 months in adults with T2D. MATERIALS AND METHODS: This retrospective cohort analysis used Optum's de-identified Market Clarity data of >79 million people to evaluate CGM use in people with T2D who were treated with non-insulin (NIT), basal insulin (BIT) and prandial insulin therapy (PIT). The primary outcomes were changes in all-cause hospitalizations, acute diabetes-related hospitalizations and acute diabetes-related emergency room visits during the 6- and 12-month post-index period following transition from blood glucose monitoring to CGM. A pre-specified subgroup analysis assessed glucose control and medication changes among people with T2D over 1 year. RESULTS: The analysis included 74 679 adults with T2D (NIT; n = 25 269), (BIT; n = 16 264) and (PIT; n = 33 146). Significant reductions in all-cause hospitalizations, acute diabetes-related hospitalizations and acute diabetes-related emergency room visits were observed in the 6-month post-index period that were sustained during the 6-12 month post-index period (NIT, -10.1%, -31.0%, -30.7%; BIT, -13.9%, -47.6%, -28.2%; and PIT, -22.6%, -52.7%, -36.6%, respectively). A subgroup analysis of 6030 people showed mean glycated haemoglobin reductions at approximately 3 months, which were also sustained throughout the post-index period: NIT, -1.1 (0.05)%; BIT, -1.1 (0.06)%; and PIT, -0.9 (0.04)%, p < 0.0001. CONCLUSIONS: CGM use in real-life across different therapeutic regimens in adults with T2D was associated with reductions in health care resource utilization with improved glucose control over 1 year.

Biosynthesis of peptide-nucleobase hybrids in ribosomal peptides.

The main biopolymers in nature are oligonucleotides and polypeptides. However, naturally occurring peptide-nucleobase hybrids are rare. Here we report the characterization of the founding member of a class of peptide-nucleobase hybrid natural products with a pyrimidone motif from a widely distributed ribosomally synthesized and post-translationally modified (RiPP) biosynthetic pathway. This pathway features two steps where a heteromeric RRE-YcaO-dehydrogenase complex catalyzes the formation of a six-membered pyrimidone ring from an asparagine residue on the precursor peptide, and an acyl esterase selectively recognizes this moiety to cleave the C-terminal follower peptide. Mechanistic studies reveal that the pyrimidone formation occurs in a substrate-assisted catalysis manner, requiring a His residue in the precursor to activate asparagine for heterocyclization. Our study expands the chemotypes of RiPP natural products and the catalytic scope of YcaO enzymes. This discovery opens avenues to create artificial biohybrid molecules that resemble both peptide and nucleobase, a modality of growing interest.

A narrative review of the CEPH-accredited bachelor's public health programs' curricula in the United States.

Background: Undergraduate programs in public health are becoming increasingly popular in the United States (US). The recent pandemic, growing climate instability, and the aging baby boomers have led to higher demands for skilled public health professionals at various levels of the workforce. This study examines the nature of courses being delivered in undergraduate public health programs across the United States. The goal is to assess domains, themes, competencies, and other specialized skills that are currently covered in these academic programs. Methodology: A search was conducted in February 2023 using the online CEPH program database to identify undergraduate public health programs in the US. In total, 86 institutions and 90 CEPH-accredited undergraduate public health programs were identified. Lists of public health courses were retrieved from each program, and a total of 2,259 unduplicated courses were extracted and analyzed. A content analysis of the extracted topics was conducted to generate 38 common themes among the courses offered. Coded course themes were mapped to the public health domains and competencies listed by the ASPPH and CEPH to evaluate the distribution of themes across course offerings. Results: Analysis of course themes found that Foundations of Public Health, Epidemiology, Public Health Management, Policy, and Leadership, Climate and Environmental Health, and Global Health Issues were the most prevalent. When course themes were mapped onto the ASPPH and CEPH domains of critical public health learning, "overview of public health" and "determinants of health" were the most populated domains. Programs had different emphases according to their approach, but overall, about two-thirds of course themes were focused on foundational and theoretical concepts of public health, and one-third were directed toward practical applications of public health concepts. Conclusion: As the demand for skilled public health workers continues to rise, programs will need to watch the skills and competencies required in the current working environment, as well as the ASPPH and CEPH criteria, and adjust their approach accordingly. Given the rapid changes in the public health landscape, schools and programs of public health should evaluate their curricula to ensure that they are meeting the needs of the workforce and the world.

Structural organization of pyruvate: ferredoxin oxidoreductase from the methanogenic archaeon Methanosarcina acetivorans.

Enzymes of the 2-oxoacid:ferredoxin oxidoreductase (OFOR) superfamily catalyze the reversible oxidation of 2-oxoacids to acyl-coenzyme A esters and carbon dioxide (CO2)using ferredoxin or flavodoxin as the redox partner. Although members of the family share primary sequence identity, a variety of domain and subunit arrangements are known. Here, we characterize the structure of a four-subunit family member: the pyruvate:ferredoxin oxidoreductase (PFOR) from the methane producing archaeon Methanosarcina acetivorans (MaPFOR). The 1.92 Å resolution crystal structure of MaPFOR shows a protein fold like those of single- or two-subunit PFORs that function in 2-oxoacid oxidation, including the location of the requisite thiamine pyrophosphate (TPP), and three [4Fe-4S] clusters. Of note, MaPFOR typically functions in the CO2 reductive direction, and structural comparisons to the pyruvate oxidizing PFORs show subtle differences in several regions of catalytical relevance. These studies provide a framework that may shed light on the biochemical mechanisms used to facilitate reductive pyruvate synthesis.

Histopathologic and Molecular Characterization of IDH-Mutant Prostatic Adenocarcinoma.

Gain-of-function isocitrate dehydrogenase (IDH) mutations are pathogenically significant in many tumor types and are actionable in cholangiocarcinoma, low-grade glioma, and acute myeloid leukemia. Rare IDH mutations have been described in prostatic adenocarcinoma (PCa). Recent publications suggested that psammomatous calcifications in PCa are associated with IDH1 mutation. In this retrospective study, we queried our institutional clinical sequencing database (Cohort 1), and previously published PCa datasets in cBioPortal (Cohort 2). Samples were stratified by oncogenic hotspot IDH mutations at IDH1 R132 and IDH2 R140/R172, and other non-hotspot IDH mutations. Seventeen (0.4%) cases were identified from 4033 PCa in Cohort 1 harboring mutually exclusive oncogenic hotspot IDH1 (N=15, one of which was subclonal) or IDH2 (N=2) mutations, and 20 (0.5%) cases had non-hotspot IDH1/2 mutations. Histologic review of 13 cases with IDH1 hotspot mutation and available material showed Grade Group 3 or higher disease. Immunohistochemistry was performed on cases with IDH1 hotspot mutation when possible, and showed AR, PSA, PSMA and NKX3.1 positive in all four cases stained. In Cohort 2, nine cases (0.3%) harboring IDH1 hotspot mutations were identified from 2749 patients, and nine cases carried non-hotspot IDH1/2 mutations. The combined cohorts of 23 PCa with clonal IDH1 hotspot mutations had no ETS fusions, SPOP hotspot mutations, somatic or germline alterations in BRCA1/2, ATM, RB1 or AR; 19 cases with successful microsatellite instability (MSI) testing were all microsatellite stable. Conversely, among 29 cases with non-hotspot IDH mutations, there were four with TMPRSS2::ERG fusion, six with SPOP hotspot mutations, and 10 with AR amplifications/hotspot mutations; eight were MSI-high. Notably, only two cases with IDH1 hotspot mutation had psammomatous calcifications. Our findings provide evidence that IDH1 hotspot mutations serve as driver alterations in this rare yet distinct molecular subset of PCa. Further studies are warranted to correlate response to androgen deprivation and IDH inhibitors.

Discontinuation of Antidepressants and the Risk of Medication Resumption among Community-Dwelling Older Adults with Depression in the US.

Antidepressants are among the most prescribed drugs in the US, but the current treatment patterns and modalities among older adults are unclear. This study assessed the patterns of discontinuation of antidepressants and the risk of medication resumption among community-dwelling older adults with depression. Using Medicare Current Beneficiary Survey (MCBS) data from 2015-2019, we identified 1084 beneficiaries with depression who newly initiated serotonergic antidepressants. The risk of medication resumption was explored using survival analysis. The median duration of continuous medication was 90 days. However, about 30% of patients had a treatment duration of 30 days or shorter, 26% for 31-90 days, 15% for 91-180 days, and 30% for 181 days or more. After adjusting for all covariates, patients with less than 30 days of continuous medication were half as likely to resume the medications compared to those with 91-180 days (HR: 0.49 (95% CI: 0.37, 0.65)). Nearly one-third of older adults used an antidepressant medication for a short duration with a lower risk of medication resumption. A shorter treatment duration without resumption might suggest over-prescription of antidepressants among community-dwelling older adults.

Silicon mediated heavy metal stress amelioration in fruit crops.

Fruit crops are essential for human nutrition and health, yet high level of heavy metal levels in soils can degrade fruit quality. These metals accumulate in plant roots and tissues due to factors like excessive fertilizer and pesticide use, poor waste management, and unscientific agricultural practices. Such accumulation can adversely affect plant growth, physiology, and yield. Consuming fruits contaminated with toxic metals poses significant health risks, including nervous system disorders and cancer. Various strategies, such as organic manuring, biomaterials, and modified cultivation practices have been widely researched to reduce heavy metal accumulation. Recently, silicon (Si) application has emerged as a promising and cost-effective solution for addressing biological and environmental challenges in food crops. Si, which can be applied to the soil, through foliar application or a combination of both, helps reduce toxic metal concentrations in soil and plants. Despite its potential, there is currently no comprehensive review that details Si's role in mitigating heavy metal stress in fruit crops. This review aims to explore the potential of Si in reducing heavy metal-induced damage in fruit crops while enhancing growth by alleviating heavy metal toxicity.

Galectin-9 Levels as a Potential Predictor of Intact HIV Reservoir Decay.

BACKGROUND: During antiretroviral therapy (ART), the HIV reservoir exhibits variability as cells with intact genomes decay faster than those with defective genomes, especially in the first years of therapy. The host factors influencing this decay are yet to be characterized. METHODS: Observational study in 74 PWH on ART, of whom 70 (94.6%) were male. We used the intact proviral DNA assay to measure intact proviruses and Luminex immunoassay to measure 32 inflammatory cytokines in plasma. Linear spline models, with a knot at seven years, evaluated the impact of baseline cytokine levels and their trajectories on intact HIV kinetics over these years. RESULTS: Baseline Gal-9 was the most predictive marker for intact HIV kinetics, with lower Gal-9 predicting faster decay over the subsequent seven years. For each 10-fold decrease in Gal-9 at baseline, there was a mean 45% (95%CI 14%-84%) greater decay of intact HIV genomes per year. Conversely, higher baseline ITAC, IL-17, and MIP-1α predicted faster intact HIV decreases. Longitudinal changes in MIP-3α and IL-6 levels strongly associated with intact HIV kinetics, with a 10-fold increase in MIP-3α and a 10-fold decrease in IL-6 associated with a a 9.5% and 10% faster decay of intact HIV genomes per year, respectively. CONCLUSION: The pronounced association between baseline Gal-9 levels and subsequent intact HIV decay suggests that strategies reducing Gal-9 levels could accelerate reservoir decay. Additionally, the correlations of MIP-3α and IL-6 with HIV kinetics indicate a broader cytokine-mediated regulatory network, hinting at multi-targeted interventions that could modulate HIV reservoir dynamics.

Maize genotypes with favourable dgat1-2 and fatb alleles possess stable high kernel oil and better fatty acid health and nutritive indices.

Diverse uses of maize oil attracted various stakeholders, including food, feed, and bioenergy, highlighting the increased demand for sustainable production. Here, 48 diverse sub-tropical maize genotypes varying for dgat1-2 and fatb genes governing oil attributes, were evaluated in three diverse locations to assess trends of oil content, fatty acid (FA) profile, the effect of environment on oil attributes, the impact of different gene combinations and determine FA health and nutritional properties. The genotypes revealed wide variation in oil content (OC: 3.4-6.8 %) and FA compositional traits, namely palmitic (PA, 11.3-24.1 %), oleic (OA, 21.5-42.7 %), linoleic (LA, 36.6-61.7 %), and linolenic (ALA, 0.7-2.3 %) acids. Double-mutants with both favourable alleles (dd/ff) exhibited 51.6 % higher oil, 33.2 % higher OA, and 30.2 % reduced PA compared to wild-types (d+d+/f+f+) across locations. These double-mutants had lower saturated FA (12.2 %), and higher unsaturated FA (87.0 %), indicating reduced susceptibility to autooxidation, with lower atherogenicity (0.14), thrombogenicity (0.27) and peroxidisability (48.15), higher cholesterolemic index (7.16), optimum oxidability (5.27) and higher nutritive-value-index (3.35) compared to d+d+/f+f+, making them promising for significant health and nutritional benefits. Locally adapted stable novel double-mutants with high-oil and better FA properties identified here can expedite the maize breeding programs, meeting production demands and addressing long-standing challenges for breeders.

Understanding the dynamics of Meloidogyne incognita infestation in pomegranate orchards of Himachal Pradesh, India (year 2018, 2019 and 2021) and its management strategies.

This study investigates the prevalence and dynamics of pomegranate wilt disease induced by Meloidogyne incognita across the Kullu, Mandi and Solan districts of Himachal Pradesh (India), revealed notable spatial and temporal variations in nematode populations and galling severity across the regions. The highest average nematode infestation of 9.25 % was observed at Nauni with highest counts of average second-stage juvenile (J2) larvae (449 larvae per 100 cc of soil) followed by Hurla (Kullu) with 7.42 % infestation. Correlation analysis reveals a strong positive relationship between larval population and galling severity suggesting a potential link between nematode levels and plant damage. Common disease symptoms were leaf size reduction, yellowing and gradual decline of pomegranate plants, often observed in patches within orchards. Microscopic identification revealed distinctive pear-shaped body of mature females while J2 larvae displayed vermiform shapes and the associated species of M. incognita was confirmed through examination of the perineal pattern. Pathogenicity test reveals initiation of leaf yellowing symptom after 45 days of inoculation of larval suspension and root galling was observed after 60 days onward followed by plant decline under greenhouse conditions. Results from pot and field experiments demonstrated the efficacy of Fluopyram and Fluensulfone in reducing nematode populations and galling severity. Treatment with drenching of Fluopyram at the rate of 2 ml/L reduced 98.56 % larvae under field and 99.00 % larvae/100 cc soil under pot conditions. Statistical analysis (paired t-test and MANOVA) confirms significant differences in galling severity and larval population before and after drenching. The study also underscores the importance of weed management in disease mitigation as several weed species (Chenopodium album and Solanum nigrum) were identified as potential reservoirs for M. incognita in infested pomegranate plant basin. This investigation contributes to the advancement of management practices for pomegranate cultivation that addresses both nematode and weed infestations ultimately enhancing crop resilience and productivity.

Long-term Outcomes of Regressed or "Burnt Out" Primary Testicular Germ Cell Tumors.

OBJECTIVE: To review the presentation and long-term oncologic outcomes of patients with regressed ("burnt out") primary testicular germ cell tumors (GCT). Certain testicular GCT can present with complete regression of the primary tumor. It is not well established if this is associated with more aggressive disease or worse oncologic outcomes. METHODS: We queried our prospectively maintained testicular cancer clinical database at a tertiary cancer center and identified patients without prior chemotherapy who had regressed primary GCT at radical orchiectomy from 1990 to 2023. All specimens were reviewed by a genitourinary pathologist at diagnosis. Long-term clinical outcomes were reported by Kaplan-Meier method. RESULTS: Fifty-six patients met inclusion criteria; at diagnosis, 17 had no evidence of extra-testicular disease and 39 had advanced (clinical stage [CS] II+) GCT. All CSx (no viable disease or germ cell neoplasia in situ at orchiectomy, and no evidence of advanced disease) and CS0 patients were managed with surveillance and had 5-year recurrence-free survival (RFS) of 88% (95% CI: 39%, 98%). All patients with CS II+ disease underwent primary treatment with surgery (n = 5) or first-line chemotherapy (n = 34). Two- and 5-year RFS for patients with CSII+ disease was 94% (95% CI: 78%, 98%) and 90% (95% CI: 72%, 97%), respectively. CONCLUSION: Patients with regressed primary testicular GCT often present with advanced disease, possibly due to lack of early clinical signs from the primary tumor. Our analysis shows excellent long-term oncologic outcomes similar to those reported in the literature for patients with viable primary testicular GCT.

Unraveling popping quality through insights on kernel physical, agro-morphological, and quality traits of diverse popcorn (Zea mays var. everta) inbreds from indigenous and exotic germplasm.

Popcorn is a specialty corn with worldwide popularity as a snack. Despite having great market demand, genetic improvement in popping quality is limited, which is caused by the limited germplasm utilization and narrow genetic base. An assortment of diverse germplasm, their effective characterization, and integration into popcorn breeding pipeline is the foundation for an efficient breeding program. Here, kernel characteristics, popping quality traits, and agro-morphological traits were evaluated across three locations on a diverse panel of 48 popcorn inbreds derived from diverse landraces and populations of exotic and indigenous origin. The variations due to genotypes, locations, and genotype × location interaction were highly significant. The popping quality traits recorded wide variation with a high coefficient of genotypic determination. The kernel dimensions, kernel density, test weight, and grain yield were negatively correlated with popping quality traits. Genotypes with rice-type kernels exhibited better popping quality than pearl-type kernels. Analysis of genotype × location (G×L) interaction identified two target locations for the key popping quality trait, popping expansion volume. PMI-PC-175, PMI-PC-187, PMI-PC-188, and PMI-PC-189 were identified as superior genotypes over checks for desirable popping quality, agronomic performance, and high grain yield. The contrasting inbreds for popping quality and flake shape (mushroom vs. butterfly) can be utilized for developing mapping populations to enhance our understanding of molecular aspects of popping quality traits. Further, the promising inbreds can be utilized in the genetic improvement of popcorn and crossed to develop superior popcorn hybrids. The results suggest a potential opportunity to establish an efficient popcorn breeding program.

Differential NEUROD1, ASCL1, and POU2F3 Expression Defines Molecular Subsets of Bladder Small Cell/Neuroendocrine Carcinoma With Prognostic Implications.

Small cell carcinomas (SMC) of the lung are now molecularly classified based on the expression of transcriptional regulators (NEUROD1, ASCL1, POU2F3, and YAP1) and DLL3, which has emerged as an investigational therapeutic target. PLCG2 has been shown to identify a distinct subpopulation of lung SMC with stem cell-like and prometastasis features and poor prognosis. We analyzed the expression of these novel neuroendocrine markers and their association with traditional neuroendocrine markers and patient outcomes in a cohort of bladder neuroendocrine carcinoma (NEC) consisting of 103 SMC and 19 large cell NEC (LCNEC) assembled in tissue microarrays. Coexpression patterns were assessed and integrated with detailed clinical annotation including overall (OS) and recurrence-free survival (RFS) and response to neoadjuvant/adjuvant chemotherapy. We identified 5 distinct molecular subtypes in bladder SMC based on the expression of ASCL1, NEUROD1, and POU2F3: ASCL1+/NEUROD1- (n = 33; 34%), ASCL1- /NEUROD1+ (n = 21; 21%), ASCL1+/NEUROD1+ (n = 17; 17%), POU2F3+ (n = 22, 22%), and ASCL1- /NEUROD1- /POU2F3- (n = 5, 5%). POU2F3+ tumors were mutually exclusive with those expressing ASCL1 and NEUROD1 and exhibited lower expression of traditional neuroendocrine markers. PLCG2 expression was noted in 33 tumors (32%) and was highly correlated with POU2F3 expression (P < .001). DLL3 expression was high in both SMC (n = 72, 82%) and LCNEC (n = 11, 85%). YAP1 expression was enriched in nonneuroendocrine components and negatively correlated with all neuroendocrine markers. In patients without metastatic disease who underwent radical cystectomy, PLCG2+ or POU2F3+ tumors had shorter RFS and OS (P < .05), but their expression was not associated with metastasis status or response to neoadjuvant/adjuvant chemotherapy. In conclusion, the NEC of the bladder can be divided into distinct molecular subtypes based on the expression of ASCL1, NEUROD1, and POU2F3. POU2F3-expressing tumors represent an ASCL1/NEUROD1-negative subset of bladder NEC characterized by lower expression of traditional neuroendocrine markers. Marker expression patterns were similar in SMC and LCNEC. Expression of PLCG2 and POU2F3 was associated with shorter RFS and OS. DLL3 was expressed at high levels in both SMC and LCNEC of the bladder, nominating it as a potential therapeutic target.

Epistasis mediates the evolution of the receptor binding mode in recent human H3N2 hemagglutinin.

The receptor-binding site of influenza A virus hemagglutinin partially overlaps with major antigenic sites and constantly evolves. In this study, we observe that mutations G186D and D190N in the hemagglutinin receptor-binding site have coevolved in two recent human H3N2 clades. X-ray crystallography results show that these mutations coordinately drive the evolution of the hemagglutinin receptor binding mode. Epistasis between G186D and D190N is further demonstrated by glycan binding and thermostability analyses. Immunization and neutralization experiments using mouse and human samples indicate that the evolution of receptor binding mode is accompanied by a change in antigenicity. Besides, combinatorial mutagenesis reveals that G186D and D190N, along with other natural mutations in recent H3N2 strains, alter the compatibility with a common egg-adaptive mutation in seasonal influenza vaccines. Overall, our findings elucidate the role of epistasis in shaping the recent evolution of human H3N2 hemagglutinin and substantiate the high evolvability of its receptor-binding mode.

G-protein-coupled receptor 84 regulates acute inflammation in normal and diabetic skin wounds.

Lipids have emerged as potent regulators of immune cell function. In the skin, adipocyte lipolysis increases the local pool of free fatty acids and is essential for coordinating early macrophage inflammation following injury. Here, we investigate G-protein-coupled receptor 84 (GPR84), a medium-chain fatty acid (MCFA) receptor, for its potential to propagate pro-inflammatory signaling after skin injury. GPR84 signaling was identified as a key component of regulating myeloid cell numbers and subsequent tissue repair through in vivo administration of a pharmacological antagonist and the MCFA decanoic acid. We found that impaired injury-induced dermal adipocyte lipolysis is a hallmark of diabetes, and lipidomic analysis demonstrated that MCFAs are significantly reduced in diabetic murine wounds. Furthermore, local administration of decanoic acid rescued myeloid cell numbers and tissue repair during diabetic wound healing. Thus, GPR84 is a readily targetable lipid signaling pathway for manipulating injury-induced tissue inflammation with beneficial effects on acute diabetic healing.

Seed endophytic bacterium Lysinibacillus sp. (ZM1) from maize (Zea mays L.) shapes its root architecture through modulation of auxin biosynthesis and nitrogen metabolism.

Seed endophytic bacteria have been shown to promote the growth and development of numerous plants. However, the underlying mechanism still needs to be better understood. The present study aims to investigate the role of a seed endophytic bacterium Lysinibacillus sp. (ZM1) in promoting plant growth and shaping the root architecture of maize seedlings. The study explores how bacteria-mediated auxin biosynthesis and nitrogen metabolism affect plant growth promotion and shape the root architecture of maize seedlings. The results demonstrate that ZM1 inoculation significantly enhances root length, root biomass, and the number of seminal roots in maize seedlings. Additionally, the treated seedlings exhibit increased shoot biomass and higher levels of photosynthetic pigments. Confocal laser scanning microscopy (CLSM) analysis revealed extensive colonization of ZM1 on root hairs, as well as in the cortical and stellar regions of the root. Furthermore, LC-MS analysis demonstrated elevated auxin content in the roots of the ZM1 treated maize seedlings compared to the uninoculated control. Inoculation with ZM1 significantly increased the levels of endogenous ammonium content, GS, and GOGAT enzyme activities in the roots of treated maize seedlings compared to the control, indicating enhanced nitrogen metabolism. Furthermore, inoculation of bacteria under nitrogen-deficient conditions enhanced plant growth, as evidenced by increased root shoot length, fresh and dry weights, average number of seminal roots, and content of photosynthetic pigments. Transcript analysis indicated upregulation of auxin biosynthetic genes, along with genes involved in nitrogen metabolism at different time points in roots of ZM1-treated maize seedlings. Collectively, our findings highlight the positive impact of Lysinibacillus sp. ZM1 inoculation on maize seeds by improving root architecture through modulation of auxin biosynthesis and affecting various nitrogen metabolism related parameters. These findings provide valuable insights into the potential utilization of seed endophytic bacteria as biofertilizers to enhance plant growth and yield in nutrient deficient soils.

Epigenetic control of multiple genes with a lentiviral vector encoding transcriptional repressors fused to compact zinc finger arrays.

Gene silencing without gene editing holds great potential for the development of safe therapeutic applications. Here, we describe a novel strategy to concomitantly repress multiple genes using zinc finger proteins fused to Krüppel-Associated Box repression domains (ZF-Rs). This was achieved via the optimization of a lentiviral system tailored for the delivery of ZF-Rs in hematopoietic cells. We showed that an optimal design of the lentiviral backbone is crucial to multiplex up to three ZF-Rs or two ZF-Rs and a chimeric antigen receptor. ZF-R expression had no impact on the integrity and functionality of transduced cells. Furthermore, gene repression in ZF-R-expressing T cells was highly efficient in vitro and in vivo during the entire monitoring period (up to 10 weeks), and it was accompanied by epigenetic remodeling events. Finally, we described an approach to improve ZF-R specificity to illustrate the path toward the generation of ZF-Rs with a safe clinical profile. In conclusion, we successfully developed an epigenetic-based cell engineering approach for concomitant modulation of multiple gene expressions that bypass the risks associated with DNA editing.