A5069125945- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- MicroRNA in disease regulation
- DNA and Nucleic Acid Chemistry
- CRISPR and Genetic Engineering
- RNA Research and Splicing
- Extracellular vesicles in disease
- RNA and protein synthesis mechanisms
- Immunotherapy and Immune Responses
- Virus-based gene therapy research
- Reproductive System and Pregnancy
- Retinal Development and Disorders
- Pregnancy and preeclampsia studies
- Cytokine Signaling Pathways and Interactions
- Prion Diseases and Protein Misfolding
- Cancer Research and Treatments
- Graphene and Nanomaterials Applications
- Lipid metabolism and biosynthesis
- Liver Disease Diagnosis and Treatment
- Neurological disorders and treatments
- Genetic Neurodegenerative Diseases
- Bacteriophages and microbial interactions
- Electrospun Nanofibers in Biomedical Applications
- melanin and skin pigmentation
- Neurogenesis and neuroplasticity mechanisms
University of Massachusetts Chan Medical School
2016-2025
UMass Memorial Medical Center
2024
University of Massachusetts Boston
2024
Massachusetts Academy of Math and Science
2024
University of Worcester
2017
Institute of Molecular Medicine
2017
Delivery represents a significant barrier to the clinical advancement of oligonucleotide therapeutics for treatment neurological disorders, such as Huntington's disease. Small, endogenous vesicles known exosomes have potential act delivery vehicles, but robust and scalable methods loading RNA therapeutic cargo into are lacking. Here, we show that hydrophobically modified small interfering RNAs (hsiRNAs) efficiently load upon co-incubation, without altering vesicle size distribution or...
Small interfering RNA (siRNA)-based drugs require chemical modifications or formulation to promote stability, minimize innate immunity, and enable delivery target tissues. Partially modified siRNAs (up 70% of the nucleotides) provide significant stabilization in vitro are commercially available; thus commonly used evaluate efficacy bio-conjugates for vivo delivery. In contrast, most clinically-advanced non-formulated compounds, using conjugation as a strategy, fully chemically (100%...
Applications of RNA interference for neuroscience research have been limited by a lack simple and efficient methods to deliver oligonucleotides primary neurons in culture the brain. Here, we show that rapidly internalize hydrophobically modified siRNAs (hsiRNAs) added directly medium without lipid formulation. We identify functional hsiRNAs targeting mRNA huntingtin, mutation which is responsible Huntington's disease, direct uptake induces potent specific silencing vitro. Moreover, single...
siRNAs are a new class of therapeutic modalities with promising clinical efficacy that requires modification or formulation for delivery to the tissue and cell interest. Conjugation lipophilic groups supports efficient cellular uptake by mechanism is not well characterized. Here we study internalization asymmetric, chemically stabilized, cholesterol-modified (sd-rxRNAs®) efficiently enter cells tissues without need formulation. We demonstrate rapid significant membrane association within...
Abstract RNA-based drugs depend on chemical modifications to increase potency and decrease immunogenicity in vivo. Chemical modification will likely improve the guide RNAs involved CRISPR-Cas9-based therapeutics as well. Cas9 orthologs are RNA-guided microbial effectors that cleave DNA. Here, we explore at all positions of crRNA tracrRNA cofactor. We identify several heavily modified versions more potent than their unmodified counterparts. In addition, describe fully chemically crRNAs...
5΄-Vinylphosphonate modification of siRNAs protects them from phosphatases, and improves silencing activity. Here, we show that 5΄-vinylphosphonate confers novel properties to siRNAs. Specifically, (i) increases siRNA accumulation in tissues, (ii) extends duration multiple organs (iii) 5΄-to-3΄ exonucleases. Delivery conjugated requires extensive chemical modifications achieve stability vivo. Because chemically modified are poor substrates for phosphorylation by kinases, 5΄-phosphate is...
Extracellular vesicles are promising delivery for therapeutic RNAs. Small interfering RNA (siRNA) conjugation to cholesterol enables efficient and reproducible loading of extracellular with the cargo. siRNAs typically chemically modified fit an application. However, siRNA chemical modification pattern has not been specifically optimized vesicle-mediated delivery. Here we used cholesterol-conjugated, hydrophobically asymmetric (hsiRNAs) evaluate effect backbone, 5'-phosphate, linker...
Abstract Inhibition of Janus kinase (JAK) family enzymes is a popular strategy for treating inflammatory and autoimmune skin diseases. In the clinic, small molecule JAK inhibitors show distinct efficacy safety profiles, likely reflecting variable selectivity subtypes. Absolute subtype has not yet been achieved. Here, we rationally design interfering RNAs (siRNAs) that offer sequence-specific gene silencing JAK1, narrowing spectrum action on JAK-dependent cytokine signaling to maintain...
Identifying therapeutic oligonucleotides that are cross-reactive to experimental animal species can dramatically accelerate the process of preclinical development and clinical translation. Here, we identify fully chemically-modified small interfering RNAs (siRNAs) Janus kinase 1 (JAK1) in humans a large variety other species. We validated identified siRNAs silencing JAK1 cell lines skin tissues multiple is one four members JAK family tyrosine kinases mediate signaling transduction many...
Abstract Small interfering RNAs are a new class of drugs, exhibiting sequence-driven, potent, and sustained silencing gene expression in vivo. We recently demonstrated that siRNA chemical architectures can be optimized to provide efficient delivery the CNS, enabling development CNS-targeted therapeutics. Many genetically-defined neurodegenerative disorders dominant, favoring selective mutant allele. In some cases, successfully targeting allele requires single nucleotide polymorphism (SNP)...
Nonalcoholic steatohepatitis (NASH) is a severe liver disorder characterized by triglyceride accumulation, inflammation, and fibrosis. With the recent increase in prevalence, NASH now leading cause of transplant, with no approved therapeutics available. Although exact molecular mechanism progression not well understood, widely held hypothesis that fat accumulation primary driver disease. Therefore, diacylglycerol O-acyltransferase 2 (DGAT2), key enzyme synthesis, has been explored as target....
Abstract Divalent short-interfering RNA (siRNA) holds promise as a therapeutic approach allowing for the sequence-specific modulation of target gene within central nervous system (CNS). However, an siRNA modality capable simultaneously modulating pairs would be invaluable treating complex neurodegenerative disorders, where more than one pathway contributes to pathogenesis. Currently, parameters and scaffold considerations multi-targeting nucleic acid modalities in CNS are undefined. Here, we...
One of the major obstacles to pharmaceutical success oligonucleotide therapeutics (ONTs) is efficient delivery from point injection intracellular setting where functional gene silencing occurs. In particular, a significant fraction internalized ONTs are nonproductively sequestered in endo-lysosomal compartments. Here, we describe two-step, robust assay for high-throughput de novo detection small bioactive molecules that enhance cellular uptake, endosomal escape, and efficacy ONTs. Using this...
Preclinical development of RNA interference (RNAi)-based therapeutics requires a rapid, accurate, and robust method simultaneously quantifying mRNA knockdown in hundreds samples. The most well-established to achieve this is quantitative real-time polymerase chain reaction (qRT-PCR), labor-intensive methodology that sample purification, which increases the potential introduce additional bias. Here, we describe QuantiGene(®) branched DNA (bDNA) assay linked 96-well Qiagen TissueLyser II quick...
We have synthesized and studied the biological biophysical properties of triazole-linked <italic>ribo</italic> <italic>xylo</italic> locked nucleic acid (LNA).
Inherited retinal dystrophies caused by dominant mutations in photoreceptor (PR) cell expressed genes are a major cause of irreversible vision loss. Oligonucleotide therapy has been interest diseases that conventional medicine cannot target. In the early days, small interfering RNAs (siRNAs) were explored clinical trials for disorders with limited success due to lack stability and efficient cellular delivery. Thus, an unmet need exists identify siRNA chemistry targets PR genes. Here, we...
Huntington's disease (HD) is caused by CAG repeat expansion within the HTT gene, with dysfunction and eventual loss of striatal medium spiny neurons a notable feature. Since receive high amounts synaptic input, we hypothesised that this vulnerability originates from an inability to sustain presynaptic performance during intense neuronal activity. To test hypothesis, primary cultures either hippocampal or were prepared wild-type mice knock-in HD mouse model which contains 140 poly-glutamine...
Effective systemic delivery of small interfering RNAs (siRNAs) to tissues other than liver remains a challenge. siRNAs are (∼15 kDa) and therefore rapidly cleared by the kidneys, resulting in limited blood residence times tissue exposure. Current strategies improve unfavorable pharmacokinetic (PK) properties rely on enhancing binding serum proteins through extensive phosphorothioate modifications or conjugation targeting ligands. Here, we describe an alternative strategy for PK based dynamic...