Antisense oligonucleotide-based therapeutic strategy for progranulin-deficient frontotemporal dementia

Loss-of-function GRN mutations result in progranulin haploinsufficiency and are a common cause of frontotemporal dementia (FTD). Antisense oligonucleotides (ASOs) are emerging as a promising therapeutic modality for neurological diseases, but ASO-based strategies for increasing target protein levels are still relatively limited. Here, we report the use of ASOs to increase progranulin protein levels by targeting the miR-29b binding site in the 3′ UTR of the GRN mRNA, resulting in increased translation.

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Antisense Oligonucleotide Therapies for Neurodegenerative Diseases

Annual Review of Neuroscience ◽

10.1146/annurev-neuro-070918-050501 ◽

2019 ◽

Vol 42 (1) ◽

pp. 385-406 ◽

Cited By ~ 41

Author(s):

C. Frank Bennett ◽

Adrian R. Krainer ◽

Don W. Cleveland

Keyword(s):

Neurodegenerative Diseases ◽

Spinal Muscular Atrophy ◽

Antisense Oligonucleotide ◽

Antisense Oligonucleotides ◽

Muscular Atrophy ◽

Neurological Diseases ◽

Great Promise ◽

Disease Symptoms ◽

Therapeutic Platform ◽

Lateral Sclerosis

Antisense oligonucleotides represent a novel therapeutic platform for the discovery of medicines that have the potential to treat most neurodegenerative diseases. Antisense drugs are currently in development for the treatment of amyotrophic lateral sclerosis, Huntington's disease, and Alzheimer's disease, and multiple research programs are underway for additional neurodegenerative diseases. One antisense drug, nusinersen, has been approved for the treatment of spinal muscular atrophy. Importantly, nusinersen improves disease symptoms when administered to symptomatic patients rather than just slowing the progression of the disease. In addition to the benefit to spinal muscular atrophy patients, there are discoveries from nusinersen that can be applied to other neurological diseases, including method of delivery, doses, tolerability of intrathecally delivered antisense drugs, and the biodistribution of intrathecal dosed antisense drugs. Based in part on the early success of nusinersen, antisense drugs hold great promise as a therapeutic platform for the treatment of neurological diseases.

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Phospholamban antisense oligonucleotides improve cardiac function in murine cardiomyopathy

Nature Communications ◽

10.1038/s41467-021-25439-0 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Niels Grote Beverborg ◽

Daniela Später ◽

Ralph Knöll ◽

Alejandro Hidalgo ◽

Steve T. Yeh ◽

...

Keyword(s):

Antisense Oligonucleotides ◽

Therapeutic Strategy ◽

Treatment Options ◽

Subcutaneous Administration ◽

Fold Increase ◽

Left Ventricular ◽

Therapeutic Modality ◽

Proof Of Concept ◽

Ventricular Contractility ◽

Novel Treatment

AbstractHeart failure (HF) is a major cause of morbidity and mortality worldwide, highlighting an urgent need for novel treatment options, despite recent improvements. Aberrant Ca2+ handling is a key feature of HF pathophysiology. Restoring the Ca2+ regulating machinery is an attractive therapeutic strategy supported by genetic and pharmacological proof of concept studies. Here, we study antisense oligonucleotides (ASOs) as a therapeutic modality, interfering with the PLN/SERCA2a interaction by targeting Pln mRNA for downregulation in the heart of murine HF models. Mice harboring the PLN R14del pathogenic variant recapitulate the human dilated cardiomyopathy (DCM) phenotype; subcutaneous administration of PLN-ASO prevents PLN protein aggregation, cardiac dysfunction, and leads to a 3-fold increase in survival rate. In another genetic DCM mouse model, unrelated to PLN (Cspr3/Mlp−/−), PLN-ASO also reverses the HF phenotype. Finally, in rats with myocardial infarction, PLN-ASO treatment prevents progression of left ventricular dilatation and improves left ventricular contractility. Thus, our data establish that antisense inhibition of PLN is an effective strategy in preclinical models of genetic cardiomyopathy as well as ischemia driven HF.

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Myoblasts and macrophages are required for therapeutic morpholino antisense oligonucleotide delivery to dystrophic muscle

Nature Communications ◽

10.1038/s41467-017-00924-7 ◽

2017 ◽

Vol 8 (1) ◽

Cited By ~ 18

Author(s):

James S. Novak ◽

Marshall W. Hogarth ◽

Jessica F. Boehler ◽

Marie Nearing ◽

Maria C. Vila ◽

...

Keyword(s):

Clinical Trials ◽

Duchenne Muscular Dystrophy ◽

Antisense Oligonucleotide ◽

Antisense Oligonucleotides ◽

Therapeutic Strategy ◽

Exon Skipping ◽

Efficient Production ◽

Dystrophic Muscle ◽

Oligonucleotide Delivery ◽

Dystrophin Protein

Abstract Exon skipping is a promising therapeutic strategy for Duchenne muscular dystrophy (DMD), employing morpholino antisense oligonucleotides (PMO-AO) to exclude disruptive exons from the mutant DMD transcript and elicit production of truncated dystrophin protein. Clinical trials for PMO show variable and sporadic dystrophin rescue. Here, we show that robust PMO uptake and efficient production of dystrophin following PMO administration coincide with areas of myofiber regeneration and inflammation. PMO localization is sustained in inflammatory foci where it enters macrophages, actively differentiating myoblasts and newly forming myotubes. We conclude that efficient PMO delivery into muscle requires two concomitant events: first, accumulation and retention of PMO within inflammatory foci associated with dystrophic lesions, and second, fusion of PMO-loaded myoblasts into repairing myofibers. Identification of these factors accounts for the variability in clinical trials and suggests strategies to improve this therapeutic approach to DMD.

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Towards a combined therapy for spinal muscular atrophy based on opposing effects of an antisense oligonucleotide on chromatin and splicing

10.1101/2021.09.24.461646 ◽

2021 ◽

Author(s):

Luciano E. Marasco ◽

Gwendal Dujardin ◽

Rui Sousa-Luís ◽

Ying Hsiu Liu ◽

José Stigliano ◽

...

Keyword(s):

Spinal Muscular Atrophy ◽

Rna Polymerase Ii ◽

Antisense Oligonucleotide ◽

Muscular Atrophy ◽

Synergistic Effects ◽

Combined Therapy ◽

Hdac Inhibitors ◽

Transcriptional Elongation ◽

Loss Of Function ◽

Protein Levels

SummarySpinal Muscular Atrophy (SMA) is a motor-neuron disease caused by loss-of-function mutations of the SMN1 gene. Humans have a paralog, SMN2, whose exon 7 is predominantly skipped, and so it cannot fully compensate for the lack of SMN1. Nusinersen (Spinraza) is a splicing-correcting antisense oligonucleotide drug (ASO) approved for clinical use. Nusinersen targets a splicing silencer located in SMN2 intron 7 pre-mRNA and, by blocking the binding of the splicing repressors hnRNPA1 and A2, it promotes higher E7 inclusion, increasing SMN protein levels. We show here that, by promoting transcriptional elongation, histone deacetylase (HDAC) inhibitors cooperate with a nusinersen-like ASO to upregulate E7 inclusion. Surprisingly, the ASO also elicits the deployment of the silencing histone mark H3K9me2 on the SMN2 gene, creating a roadblock to RNA polymerase II elongation that acts negatively on E7 inclusion. By removing the roadblock, HDAC inhibition counteracts the undesired chromatin effects of the ASO, resulting in higher E7 inclusion. Combined systemic administration of the nusinersen-like ASO and HDAC inhibitors in neonate SMA mice had strong synergistic effects on SMN expression, growth, survival, and neuromuscular function. Thus, we suggest that HDAC inhibitors have the potential to increase the clinical efficacy of nusinersen, and perhaps other splicing-modulatory ASO drugs, without large pleiotropic effects, as assessed by genome-wide analyses.

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Treponema, Iron and Neurodegeneration

Current Alzheimer Research ◽

10.2174/1567205013666161122093404 ◽

2018 ◽

Vol 15 (8) ◽

pp. 716-722 ◽

Cited By ~ 1

Author(s):

A. Jolivet-Gougeon ◽

M. Bonnaure-Mallet

Keyword(s):

Iron Metabolism ◽

Plasma Proteins ◽

Iron Homeostasis ◽

Neurological Diseases ◽

Host Cells ◽

Loss Of Function ◽

Binding Ability ◽

Iron Binding Proteins ◽

Oxidative Species ◽

Iron Reductase

Spirochetes are suspected to be linked to the genesis of neurological diseases, including neurosyphillis or neurodegeneration (ND). Impaired iron homeostasis has been implicated in loss of function in several enzymes requiring iron as a cofactor, formation of toxic oxidative species, inflammation and elevated production of beta-amyloid proteins. This review proposes to discuss the link that may exist between the involvement of Treponema spp. in the genesis or worsening of ND, and iron dyshomeostasis. Proteins secreted by Treponema can act directly on iron metabolism, with hemin binding ability (HbpA and HbpB) and iron reductase able to reduce the central ferric iron of hemin, iron-containing proteins (rubredoxin, neelaredoxin, desulfoferrodoxin metalloproteins, bacterioferritins etc). Treponema can also interact with cellular compounds, especially plasma proteins involved in iron metabolism, contributing to the virulence of the syphilis spirochetes (e.g. treponemal motility and survival). Fibronectin, transferrin and lactoferrin were also shown to be receptors for treponemal adherence to host cells and extracellular matrix. Association between Treponema and iron binding proteins results in iron accumulation and sequestration by Treponema from host macromolecules during systemic and mucosal infections.

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LOXL1 confers antiapoptosis and promotes gliomagenesis through stabilizing BAG2

Cell Death and Differentiation ◽

10.1038/s41418-020-0558-4 ◽

2020 ◽

Vol 27 (11) ◽

pp. 3021-3036 ◽

Cited By ~ 2

Author(s):

Hua Yu ◽

Jun Ding ◽

Hongwen Zhu ◽

Yao Jing ◽

Hu Zhou ◽

...

Keyword(s):

Molecular Chaperone ◽

Glioma Cell ◽

Lysyl Oxidase ◽

The Cancer Genome Atlas ◽

Loss Of Function ◽

Protein Levels ◽

Important Mediator ◽

Cancer Genome Atlas ◽

Potential Strategy ◽

Glioma Progression

Abstract The lysyl oxidase (LOX) family is closely related to the progression of glioma. To ensure the clinical significance of LOX family in glioma, The Cancer Genome Atlas (TCGA) database was mined and the analysis indicated that higher LOXL1 expression was correlated with more malignant glioma progression. The functions of LOXL1 in promoting glioma cell survival and inhibiting apoptosis were studied by gain- and loss-of-function experiments in cells and animals. LOXL1 was found to exhibit antiapoptotic activity by interacting with multiple antiapoptosis modulators, especially BAG family molecular chaperone regulator 2 (BAG2). LOXL1-D515 interacted with BAG2-K186 through a hydrogen bond, and its lysyl oxidase activity prevented BAG2 degradation by competing with K186 ubiquitylation. Then, we discovered that LOXL1 expression was specifically upregulated through the VEGFR-Src-CEBPA axis. Clinically, the patients with higher LOXL1 levels in their blood had much more abundant BAG2 protein levels in glioma tissues. Conclusively, LOXL1 functions as an important mediator that increases the antiapoptotic capacity of tumor cells, and approaches targeting LOXL1 represent a potential strategy for treating glioma. In addition, blood LOXL1 levels can be used as a biomarker to monitor glioma progression.

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Cockayne syndrome B protein acts as an ATP-dependent processivity factor that helps RNA polymerase II overcome nucleosome barriers

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2013379117 ◽

2020 ◽

Vol 117 (41) ◽

pp. 25486-25493 ◽

Cited By ~ 1

Author(s):

Jun Xu ◽

Wei Wang ◽

Liang Xu ◽

Jia-Yu Chen ◽

Jenny Chong ◽

...

Keyword(s):

Rna Polymerase Ii ◽

Neurological Diseases ◽

Transcription Elongation ◽

Cockayne Syndrome ◽

Stable Complex ◽

Loss Of Function ◽

Pol Ii ◽

Chromatin Remodelers ◽

Processivity Factor

While loss-of-function mutations in Cockayne syndrome group B protein (CSB) cause neurological diseases, this unique member of the SWI2/SNF2 family of chromatin remodelers has been broadly implicated in transcription elongation and transcription-coupled DNA damage repair, yet its mechanism remains largely elusive. Here, we use a reconstituted in vitro transcription system with purified polymerase II (Pol II) and Rad26, a yeast ortholog of CSB, to study the role of CSB in transcription elongation through nucleosome barriers. We show that CSB forms a stable complex with Pol II and acts as an ATP-dependent processivity factor that helps Pol II across a nucleosome barrier. This noncanonical mechanism is distinct from the canonical modes of chromatin remodelers that directly engage and remodel nucleosomes or transcription elongation factors that facilitate Pol II nucleosome bypass without hydrolyzing ATP. We propose a model where CSB facilitates gene expression by helping Pol II bypass chromatin obstacles while maintaining their structures.

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TRIM27 interacts with Iκbα to promote the growth of human renal cancer cells through regulating the NF-κB pathway

BMC Cancer ◽

10.1186/s12885-021-08562-5 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Chengwu Xiao ◽

Wei Zhang ◽

Meimian Hua ◽

Huan Chen ◽

Bin Yang ◽

...

Keyword(s):

Cell Lines ◽

Prognostic Indicator ◽

The Cancer Genome Atlas ◽

Mrna Levels ◽

Loss Of Function ◽

Protein Levels ◽

Kaplan Meier ◽

Cancer Genome Atlas

Abstract Background The tripartite motif (TRIM) family proteins exhibit oncogenic roles in various cancers. The roles of TRIM27, a member of the TRIM super family, in renal cell carcinoma (RCC) remained unexplored. In the current study, we aimed to investigate the clinical impact and roles of TRIM27 in the development of RCC. Methods The mRNA levels of TRIM27 and Kaplan–Meier survival of RCC were analyzed from The Cancer Genome Atlas database. Real-time PCR and Western blotting were used to measure the mRNA and protein levels of TRIM27 both in vivo and in vitro. siRNA and TRIM27 were exogenously overexpressed in RCC cell lines to manipulate TRIM27 expression. Results We discovered that TRIM27 was elevated in RCC patients, and the expression of TRIM27 was closely correlated with poor prognosis. The loss of function and gain of function results illustrated that TRIM27 promotes cell proliferation and inhibits apoptosis in RCC cell lines. Furthermore, TRIM27 expression was positively associated with NF-κB expression in patients with RCC. Blocking the activity of NF-κB attenuated the TRIM27-mediated enhancement of proliferation and inhibition of apoptosis. TRIM27 directly interacted with Iκbα, an inhibitor of NF-κB, to promote its ubiquitination, and the inhibitory effects of TRIM27 on Iκbα led to NF-κB activation. Conclusions Our results suggest that TRIM27 exhibits an oncogenic role in RCC by regulating NF-κB signaling. TRIM27 serves as a specific prognostic indicator for RCC, and strategies targeting the suppression of TRIM27 function may shed light on future therapeutic approaches.

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Cerebrospinal Fluid Parameters in Antisense Oligonucleotide-Treated Adult 5q-Spinal Muscular Atrophy Patients

Brain Sciences ◽

10.3390/brainsci11030296 ◽

2021 ◽

Vol 11 (3) ◽

pp. 296

Author(s):

Lars Hendrik Müschen ◽

Alma Osmanovic ◽

Camilla Binz ◽

Konstantin F. Jendretzky ◽

Gresa Ranxha ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Spinal Muscular Atrophy ◽

Total Protein ◽

Antisense Oligonucleotide ◽

Muscular Atrophy ◽

Neurological Diseases ◽

Clinical Signs ◽

Central Nervous System Disease ◽

Oligoclonal Bands ◽

System Disease

Approval of nusinersen, an intrathecally administered antisense oligonucleotide, for the treatment of 5q-spinal muscular atrophy (SMA) marked the beginning of a new therapeutic era in neurological diseases. Changes in routine cerebrospinal fluid (CSF) parameters under nusinersen have only recently been described in adult SMA patients. We aimed to explore these findings in a real-world setting and to identify clinical and procedure-associated features that might impact CSF parameters. Routinely collected CSF parameters (leukocyte count, lactate, total protein, CSF/serum albumin quotient (QAlbumin), oligoclonal bands) of 28 adult SMA patients were examined for up to 22 months of nusinersen treatment. Total protein and QAlbumin values significantly increased in the first 10 months, independent of the administration procedure. By month 14, no further increases were detected. Two patients developed transient pleocytosis. In two cases, positive oligoclonal bands were found in the beginning and in four patients throughout the whole observation period. No clinical signs of inflammatory central nervous system disease were apparent. Our data confirm elevated CSF total protein and QAlbumin during nusinersen treatment. These alterations may be caused by both repeated lumbar punctures and the interval between procedures rather than by the medication itself. Generally, there were no severe alterations of CSF routine parameters. These results further underline the safety of nusinersen therapy.

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Therapeutic manipulation of IKBKAP mis-splicing with a small molecule to cure familial dysautonomia

Nature Communications ◽

10.1038/s41467-021-24705-5 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Masahiko Ajiro ◽

Tomonari Awaya ◽

Young Jin Kim ◽

Kei Iida ◽

Masatsugu Denawa ◽

...

Keyword(s):

Small Molecule ◽

Genetic Disease ◽

Therapeutic Strategy ◽

Therapeutic Potential ◽

Familial Dysautonomia ◽

Therapeutic Modality ◽

Aberrant Splicing ◽

Intronic Splicing Enhancer ◽

Exon 20 ◽

Splicing Enhancer

AbstractApproximately half of genetic disease-associated mutations cause aberrant splicing. However, a widely applicable therapeutic strategy to splicing diseases is yet to be developed. Here, we analyze the mechanism whereby IKBKAP-familial dysautonomia (FD) exon 20 inclusion is specifically promoted by a small molecule splice modulator, RECTAS, even though IKBKAP-FD exon 20 has a suboptimal 5′ splice site due to the IVS20 + 6 T > C mutation. Knockdown experiments reveal that exon 20 inclusion is suppressed in the absence of serine/arginine-rich splicing factor 6 (SRSF6) binding to an intronic splicing enhancer in intron 20. We show that RECTAS directly interacts with CDC-like kinases (CLKs) and enhances SRSF6 phosphorylation. Consistently, exon 20 splicing is bidirectionally manipulated by targeting cellular CLK activity with RECTAS versus CLK inhibitors. The therapeutic potential of RECTAS is validated in multiple FD disease models. Our study indicates that small synthetic molecules affecting phosphorylation state of SRSFs is available as a new therapeutic modality for mechanism-oriented precision medicine of splicing diseases.

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