Genome editing therapies hold great promise for the cure of monogenic and other diseases; however, the application of nonviral gene delivery methods is limited by both a lack of fundamental knowledge ...of interactions of the gene-carrier in complex animals and biocompatibility. Herein, we characterize nonviral gene delivery vehicle formulations that are based on diblock polycations containing a hydrophilic and neutral glucose block chain extended with cationic secondary amines of three lengths, poly(methacrylamido glucopyranose-block-2-methylaminoethyl methacrylate) P(MAG-b-MAEMt)-1, -2, -3. These polymers were formulated with plasmid DNA to prepare polyelectrolyte complexes (polyplexes). In addition, two controls, P(EG-b-MAEMt) and P(MAEMt), were synthesized, formulated into polyplexes and the ex vivo hemocompatibility, or blood compatibility, and in vivo biodistribution of the formulations were compared to the glycopolymers. While both polymer structure and N/P (amine to phosphate) ratio were important factors affecting hemocompatibility, N/P ratio played a stronger role in determining polyplex biodistribution. P(EG-b-MAEMt) and P(MAEMt) lysed red blood cells at both high and low N/P formulations while P(MAG-b-MAEMt) did not significantly lyse cells at either formulation at short and medium polymer lengths. Conversely, P(MAG-b-MAEMt) did not affect coagulation at N/P = 5, but significantly delayed coagulation at N/P = 15. P(EG-b-MAEMt) and P(MAEMt) did not affect coagulation at either formulation. After polymer and pDNA cargo distribution was observed in vivo, P(EG-b-MAEMt) N/P = 5 and P(MAG-b-MAEMt) N/P = 5 both dissociated and deposited polymer in the liver, while pDNA cargo from P(MAG-b-MAEMt) N/P = 15 was found in the liver, lungs, and spleen. The contrast between P(MAG-b-MAEMt) at N/P = 5 and 15 demonstrates that polyplex stability in the blood can be improved with N/P ratio and potentially aid polyplex biodistribution through simply varying the formulation ratios.
Natural product feedstocks such as carbohydrates and vegetable oils offer tremendous potential for creating sustainable cross-linked epoxy resin thermosets for numerous applications. Herein, we ...designed and synthesized trehalose- and β-cyclodextrin-based carboxylic acid hardeners to cure with epoxidized soybean oil forming predominantly sustainable epoxy resins. Trehalose (Tr) and β-cyclodextrin (Cd) were functionalized with heptanoyl chloride (H) and succinic anhydride (S). The resulting carboxylic acid hardeners were homogeneously formulated and cross-linked with epoxidized soy bean oil (ESO) at three different COOH/epoxide ratios. The cured resins were thermally stable up to 300 °C and stable in neutral and acidic aqueous conditions. Yet, degradation into water-soluble components could be triggered upon exposure to basic aqueous media. The physical properties of these materials are tunable based on feedstock composition and identity of the carbohydrate hardener. The glass transition temperatures ( T g) of the Tr-based epoxy polymers ranged from −3 to 3 °C, whereas the Cd-based polymers exhibited T g values of 28–36 °C. The mechanical properties including tensile strength and Young’s moduli also varied where the Cd-thermosets offered higher performance due to the structural rigidity of the cup-like structure. Homogeneous epoxy resin films of these materials were examined for their ability to promote cell adhesion and proliferation using neonatal human dermal fibroblast (HDFn) cells. The results indicated that films composed of the Cd-based epoxy resin with a 50/50 ratio of −COOH/epoxide promoted cell adhesion and proliferation with density similar to that of the well-studied control polymer poly(d l-lactide-co-glycolide) (PLG). Interestingly, the Tr-based epoxy films completely prevented cell adhesion and growth. The starkly different cell adhesion results and favorable physical characteristics of these predominantly sustainable epoxy resins support their promise as benign surfaces and scaffolds for a variety of applications ranging from adhesives and antifouling coatings to wound healing and tissue engineering.
A series of nine poly(2-deoxy-2-methacrylamido glucopyranose)-b-poly(methacrylate amine) diblock copolycations has been synthesized as new colloidally stable polynucleotide vehicles. The cationic ...block was varied in length and in the degree of methyl group substitution (secondary, tertiary, quaternary) on the pendant amine in an effort to optimize the structure and activity for plasmid DNA (pDNA) delivery. Upon a thorough kinetic study of polymerization for each polymer, the glycopolymers were prepared with well-controlled M n and Đ. The binding and colloidal stability of the polymer–pDNA nanocomplexes at different N/P ratios and in biological media have been investigated using gel electrophoresis and light scattering techniques. The toxicity and transfection efficiency of the polyplexes have been evaluated with Hep G2 (human liver hepatocellular carcinoma) cells; several polymers displayed excellent delivery and toxicity profiles justifying their further development for in vivo gene therapy.
Mammalian gestation and pregnancy are fast evolving processes that involve the interaction of the fetal, maternal and paternal genomes. Version 1.0 of the GEneSTATION database ...(http://genestation.org) integrates diverse types of omics data across mammals to advance understanding of the genetic basis of gestation and pregnancy-associated phenotypes and to accelerate the translation of discoveries from model organisms to humans. GEneSTATION is built using tools from the Generic Model Organism Database project, including the biology-aware database CHADO, new tools for rapid data integration, and algorithms that streamline synthesis and user access. GEneSTATION contains curated life history information on pregnancy and reproduction from 23 high-quality mammalian genomes. For every human gene, GEneSTATION contains diverse evolutionary (e.g. gene age, population genetic and molecular evolutionary statistics), organismal (e.g. tissue-specific gene and protein expression, differential gene expression, disease phenotype), and molecular data types (e.g. Gene Ontology Annotation, protein interactions), as well as links to many general (e.g. Entrez, PubMed) and pregnancy disease-specific (e.g. PTBgene, dbPTB) databases. By facilitating the synthesis of diverse functional and evolutionary data in pregnancy-associated tissues and phenotypes and enabling their quick, intuitive, accurate and customized meta-analysis, GEneSTATION provides a novel platform for comprehensive investigation of the function and evolution of mammalian pregnancy.
An anatomic transcriptional atlas of human glioblastoma Puchalski, Ralph B; Shah, Nameeta; Miller, Jeremy ...
Science (American Association for the Advancement of Science),
05/2018, Letnik:
360, Številka:
6389
Journal Article
Recenzirano
Odprti dostop
Glioblastoma is an aggressive brain tumor that carries a poor prognosis. The tumor's molecular and cellular landscapes are complex, and their relationships to histologic features routinely used for ...diagnosis are unclear. We present the Ivy Glioblastoma Atlas, an anatomically based transcriptional atlas of human glioblastoma that aligns individual histologic features with genomic alterations and gene expression patterns, thus assigning molecular information to the most important morphologic hallmarks of the tumor. The atlas and its clinical and genomic database are freely accessible online data resources that will serve as a valuable platform for future investigations of glioblastoma pathogenesis, diagnosis, and treatment.
A series of nine poly(2-deoxy-2-methacrylamido glucopyranose)-
-poly(methacrylate amine) diblock copolycations The cationic block was varied in length and in the degree of methyl group substitution ...(secondary, tertiary, quaternary) on the pendant amine in an effort to optimize the structure and activity for plasmid DNA delivery. Upon a thorough kinetic study of polymerization for each polymer, the glycopolymers were prepared with well-controlled M
and Ð. The binding and colloidal stability of the polymer-pDNA nanocomplexes at different N/P ratios and in biological media has been investigated using gel electrophoresis and light scattering techniques. The toxicity and transfection efficiency of the polyplexes has been evaluated with Hep G2 (human liver hepatocellular carcinoma) cells; several polymers displayed excellent delivery and toxicity profiles justifying their further development for
gene therapy.
Dysregulation of apoptotic machinery is one mechanism by which acute myeloid leukemia (AML) acquires a clonal survival advantage. B-cell lymphoma protein-2 (BCL2) overexpression is a common feature ...in hematologic malignancies. The selective BCL2 inhibitor, venetoclax (VEN) is used in combination with azacitidine (AZA), a DNAmethyltransferase inhibitor (DNMTi), to treat patients with AML. Despite promising response rates to VEN/AZA, resistance to the agent is common. One identified mechanism of resistance is the upregulation of myeloid cell leukemia-1 protein (MCL1). Pevonedistat (PEV), a novel agent that inhibits NEDD8-activating enzyme, and AZA both upregulate NOXA (PMAIP1), a BCL2 family protein that competes with effector molecules at the BH3 binding site of MCL1. We demonstrate that PEV/AZA combination induces NOXA to a greater degree than either PEV or AZA alone, which enhances VEN-mediated apoptosis. Herein, using AML cell lines and primary AML patient samples ex vivo, including in cells with genetic alterations linked to treatment resistance, we demonstrate robust activity of the PEV/VEN/AZA triplet. These findings were corroborated in preclinical systemic engrafted models of AML. Collectively, these results provide rational for combining PEV/VEN/AZA as a novel therapeutic approach in overcoming AML resistance in current therapies.
Objective:New antidepressant treatments are needed that are effective, rapid acting, safe, and tolerable. Intermittent theta-burst stimulation (iTBS) is a noninvasive brain stimulation treatment that ...has been approved by the U.S. Food and Drug Administration for treatment-resistant depression. Recent methodological advances suggest that the current iTBS protocol might be improved through 1) treating patients with multiple sessions per day at optimally spaced intervals, 2) applying a higher overall pulse dose of stimulation, and 3) precision targeting of the left dorsolateral prefrontal cortex (DLPFC) to subgenual anterior cingulate cortex (sgACC) circuit. The authors examined the feasibility, tolerability, and preliminary efficacy of Stanford Accelerated Intelligent Neuromodulation Therapy (SAINT), an accelerated, high-dose resting-state functional connectivity MRI (fcMRI)–guided iTBS protocol for treatment-resistant depression.Methods:Twenty-two participants with treatment-resistant depression received open-label SAINT. fcMRI was used to individually target the region of the left DLPFC most anticorrelated with sgACC in each participant. Fifty iTBS sessions (1,800 pulses per session, 50-minute intersession interval) were delivered as 10 daily sessions over 5 consecutive days at 90% resting motor threshold (adjusted for cortical depth). Neuropsychological testing was conducted before and after SAINT.Results:One participant withdrew, leaving a sample size of 21. Nineteen of 21 participants (90.5%) met remission criteria (defined as a score <11 on the Montgomery-Åsberg Depression Rating Scale). In the intent-to-treat analysis, 19 of 22 participants (86.4%) met remission criteria. Neuropsychological testing demonstrated no negative cognitive side effects.Conclusions:SAINT, an accelerated, high-dose, iTBS protocol with fcMRI-guided targeting, was well tolerated and safe. Double-blinded sham-controlled trials are needed to confirm the remission rate observed in this initial study.
Abstract
Study Objectives
Cardiovascular autonomic dysfunction, as measured by short-term diurnal heart rate variability (HRV), has been reported in older adults with mild cognitive impairment (MCI). ...However, it is unclear whether this impairment also exists during sleep in this group. We, therefore, compared overnight HRV during sleep in older adults with MCI and those with subjective cognitive impairment (SCI).
Methods
Older adults (n = 210) underwent overnight polysomnography. Eligible participants were characterized as multi-domain MCI or SCI. The multi-domain MCI group was comprised of amnestic and non-amnestic subtypes. Power spectral analysis of HRV was conducted on the overnight electrocardiogram during non-rapid eye movement (NREM), rapid eye movement (REM), N1, N2, N3 sleep stages, and wake periods. High-frequency HRV (HF-HRV) was employed as the primary measure to estimate parasympathetic function.
Results
The MCI group showed reduced HF-HRV during NREM sleep (p = 0.018), but not during wake or REM sleep (p > 0.05) compared to the SCI group. Participants with aMCI compared to SCI had the most pronounced reduction in HF-HRV across all NREM sleep stages—N1, N2, and N3, but not during wake or REM sleep. The naMCI sub-group did not show any significant differences in HF-HRV during any sleep stage compared to SCI.
Conclusions
Our study showed that amnestic MCI participants had greater reductions in HF-HRV during NREM sleep, relative to those with SCI, suggesting potential vulnerability to sleep-related parasympathetic dysfunction. HF-HRV, especially during NREM sleep, may be an early biomarker for dementia detection.
CONTEXT Elucidation of the ischemic cascade has helped stimulate development
of neuroprotective drugs aimed at limiting brain injury in the hours following
an ischemic stroke. To date, none of these ...drugs has shown clinical efficacy. OBJECTIVE To examine the efficacy of gavestinel (GV150526), an antagonist of the
glycine site of the N-methyl-D-aspartate receptor,
as a neuroprotective therapy for acute ischemic stroke when administered within
6 hours of symptom onset. DESIGN The Glycine Antagonist in Neuroprotection (GAIN) Americas trial, a randomized,
double-blind placebo-controlled trial with enrollment from April 1998 to October
1999. SETTING One hundred thirty-two hospital centers across the United States and
Canada. PATIENTS The primary efficacy population consisted of 1367 ischemic stroke patients
with a predefined level of limb weakness and functional independence prior
to stroke, stratified at randomization by age (≤75 vs >75 years) and initial
stroke severity (National Institutes of Health NIH Stroke Scale scores of
2-5, 6-13, or ≥14). INTERVENTION Patients were randomly assigned to receive an intravenous loading dose
(800 mg) plus 5 maintenance doses (200 mg every 12 hours) of gavestinel (n
= 701) or placebo (n = 666) for 3 days. MAIN OUTCOME MEASURE Functional capability at 3 months, measured by the Barthel Index (BI),
with scores trichotomized as dead/0-55, 60-90, and 95-100, compared between
the gavestinel and placebo groups. RESULTS Treatment groups were well matched for baseline characteristics. For
each group, median NIH Stroke Scale was 12, median age was 72 years, and median
time to treatment was 5.2 hours. No statistically significant improvement
on the 3-month BI trichotomy was demonstrated for gavestinel (P = .79). The proportion who were functionally independent (BI score
= 95-100) was 39% in the gavestinel group and 37% in the placebo group. No
statistically significant difference in 3-month survival was observed using
Kaplan-Meier curves (P = .11). No other secondary
end point suggested an advantage for gavestinel. Among the 333 patients (24%)
who received recombinant tissue-type plasminogen activator, there was also
no benefit for gavestinel (P = .53). There were no
serious safety issues. CONCLUSION In this study, gavestinel administered up to 6 hours after an acute
ischemic stroke did not improve functional outcome at 3 months.
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