Manuel Daldin
Latina, Lazio, Italia
498 follower
497 collegamenti
Attività
-
Thrilled to announce that I’ll be representing IRBM at The Forum Community Event next Wednesday! I’m eager to discuss cutting-edge innovations in…
Thrilled to announce that I’ll be representing IRBM at The Forum Community Event next Wednesday! I’m eager to discuss cutting-edge innovations in…
Consigliato da Manuel Daldin
-
RESEARCH UPDATE - Prilenia provided an update on their filing with the European Medicines Agency (EMA). Full community letter and link to press…
RESEARCH UPDATE - Prilenia provided an update on their filing with the European Medicines Agency (EMA). Full community letter and link to press…
Consigliato da Manuel Daldin
-
🤲 The International Huntington Association is “hopeful that we get closer to a cure for Huntington’s Disease and is keen on all the research…
🤲 The International Huntington Association is “hopeful that we get closer to a cure for Huntington’s Disease and is keen on all the research…
Consigliato da Manuel Daldin
Esperienza
Formazione
-
-
Attività e associazioni:Study of defense responses of model plant Arabidopsis thaliana, particularly of signaling components endogenous elicitors mediated.
Master’s dissertation title: “Role of PCaP1, Ca-binding and membrane associated protein,
in oligogalacturonides mediated signaling in Arabidopsis thaliana”. -
Pubblicazioni
-
Identification and Optimization of RNA-Splicing Modulators as Huntingtin Protein-Lowering Agents for the Treatment of Huntington’s Disease
Journal of Medicinal Chemistry
Huntington’s disease (HD) is caused by an expanded CAG trinucleotide repeat in exon 1 of the huntingtin (HTT) gene. We report the design of a series of HTT pre-mRNA splicing modulators that lower huntingtin (HTT) protein, including the toxic mutant huntingtin (mHTT), by promoting insertion of a pseudoexon containing a premature termination codon at the exon 49−50 junction. The resulting transcript undergoes nonsense-mediated decay, leading to a reduction of HTT mRNA transcripts and protein…
Huntington’s disease (HD) is caused by an expanded CAG trinucleotide repeat in exon 1 of the huntingtin (HTT) gene. We report the design of a series of HTT pre-mRNA splicing modulators that lower huntingtin (HTT) protein, including the toxic mutant huntingtin (mHTT), by promoting insertion of a pseudoexon containing a premature termination codon at the exon 49−50 junction. The resulting transcript undergoes nonsense-mediated decay, leading to a reduction of HTT mRNA transcripts and protein levels. The starting benzamide core was modified to pyrazine amide and further optimized to give a potent, CNSpenetrant, and orally bioavailable HTT-splicing modulator 27. This compound reduced canonical splicing of the HTT RNA exon 49−50 and demonstrated significant HTT-lowering in both human HD stem cells and mouse BACHD models. Compound 27 is a structurally diverse HTTsplicing modulator that may help understand the mechanism of adverse effects such as peripheral neuropathy associated with branaplam.
-
Salivary Huntingtin protein is uniquely associated with clinical features of Huntington's disease
Scientific Reports
Measuring Huntingtin (HTT) protein in peripheral cells represents an essential step in biomarker discovery for Huntington's Disease (HD), however to date, investigations into the salivary expression of HTT has been lacking. In the current study, we quantified total HTT (tHTT) and mutant HTT (mHTT) protein in matched blood and saliva samples using single molecule counting (SMC) immunoassays: 2B7-D7F7 (tHTT) and 2B7-MW1 (mHTT). Matched samples, and clinical data, were collected from 95 subjects:…
Measuring Huntingtin (HTT) protein in peripheral cells represents an essential step in biomarker discovery for Huntington's Disease (HD), however to date, investigations into the salivary expression of HTT has been lacking. In the current study, we quantified total HTT (tHTT) and mutant HTT (mHTT) protein in matched blood and saliva samples using single molecule counting (SMC) immunoassays: 2B7-D7F7 (tHTT) and 2B7-MW1 (mHTT). Matched samples, and clinical data, were collected from 95 subjects: n = 19 manifest HD, n = 34 premanifest HD (PM), and n = 42 normal controls (NC). Total HTT and mHTT levels were not correlated in blood and saliva. Plasma tHTT was significantly associated with age, and participant sex; whereas salivary mHTT was significantly correlated with age, CAG repeat length and CAP score. Plasma and salivary tHTT did not differ across cohorts. Salivary and plasma mHTT were significantly increased in PM compared to NC; salivary mHTT was also significantly increased in HD compared to NC. Only salivary tHTT and mHTT were significantly correlated with clinical measures. Salivary HTT is uniquely associated with clinical measures of HD and offers significant promise as a relevant, non-invasive HD biomarker. Its use could be immediately implemented into both translational and clinical research applications.
-
Quantifying Huntingtin Protein in Human Cerebrospinal Fluid Using a Novel Polyglutamine Length-Independent Assay
Journal of Huntington's Disease
A novel ultrasensitive SMC immunoassay was developed to quantify HTT protein in a polyglutamine length-independent manner and shown to measure HTT in both control and HD participant CSF samples. We validate the selectivity and specificity of the readout using biochemical and molecular biology tools, and we undertook a preliminary analytical qualification of this assay to enable its clinical use. We also used this novel assay, along with the previously described mHTT assay, to analyze CSF from…
A novel ultrasensitive SMC immunoassay was developed to quantify HTT protein in a polyglutamine length-independent manner and shown to measure HTT in both control and HD participant CSF samples. We validate the selectivity and specificity of the readout using biochemical and molecular biology tools, and we undertook a preliminary analytical qualification of this assay to enable its clinical use. We also used this novel assay, along with the previously described mHTT assay, to analyze CSF from control and HD participants. The results of this preliminary set suggests that correlation is present between mHTT and the polyglutamine length-independent HTT levels in human CSF.
-
Analysis of mutant and total huntingtin expression in Huntington's disease murine models
Scientific Reports
Huntington's disease (HD) is a monogenetic neurodegenerative disorder that is caused by the expansion of a polyglutamine region within the huntingtin (HTT) protein, but there is still an incomplete understanding of the molecular mechanisms that drive pathology. Expression of the mutant form of HTT is a key aspect of diseased tissues, and the most promising therapeutic approaches aim to lower expanded HTT levels. Consequently, the investigation of HTT expression in time and in multiple tissues…
Huntington's disease (HD) is a monogenetic neurodegenerative disorder that is caused by the expansion of a polyglutamine region within the huntingtin (HTT) protein, but there is still an incomplete understanding of the molecular mechanisms that drive pathology. Expression of the mutant form of HTT is a key aspect of diseased tissues, and the most promising therapeutic approaches aim to lower expanded HTT levels. Consequently, the investigation of HTT expression in time and in multiple tissues, with assays that accurately quantify expanded and non-expanded HTT, are required to delineate HTT homeostasis and to best design and interpret pharmacodynamic readouts for HTT lowering therapeutics. Here we evaluate mutant polyglutamine-expanded (mHTT) and polyglutamine-independent HTT specific immunoassays for validation in human HD and control fibroblasts and use to elucidate the CSF/brain and peripheral tissue expression of HTT in preclinical HD models.
-
Validation of Ultrasensitive Mutant Huntingtin Detection in Human Cerebrospinal Fluid by Single Molecule Counting Immunoassay.
Journal of Huntington's Disease
BACKGROUND:
The measurement of disease-relevant biomarkers has become a major component of clinical trial design, but in the absence of rigorous clinical and analytical validation of detection methodology, interpretation of results may be misleading. In Huntington's disease (HD), measurement of the concentration of mutant huntingtin protein (mHTT) in cerebrospinal fluid (CSF) of patients may serve as both a disease progression biomarker and a pharmacodynamic readout for HTT-lowering…BACKGROUND:
The measurement of disease-relevant biomarkers has become a major component of clinical trial design, but in the absence of rigorous clinical and analytical validation of detection methodology, interpretation of results may be misleading. In Huntington's disease (HD), measurement of the concentration of mutant huntingtin protein (mHTT) in cerebrospinal fluid (CSF) of patients may serve as both a disease progression biomarker and a pharmacodynamic readout for HTT-lowering therapeutic approaches. We recently published the quantification of mHTT levels in HD patient CSF by a novel ultrasensitive immunoassay-based technology and here analytically validate it for use.
OBJECTIVE:
This work aims to analytically and clinically validate our ultrasensitive assay for mHTT measurement in human HD CSF, for application as a pharmacodynamic biomarker of CNS mHTT lowering in clinical trials.
METHODS:
The single molecule counting (SMC) assay is an ultrasensitive bead-based immunoassay where upon specific recognition, dye-labeled antibodies are excited by a confocal laser and emit fluorescent light as a readout. The detection of mHTT by this technology was clinically validated following established Food and Drug Administration and European Medicine Agency guidelines.
RESULTS:
The SMC assay was demonstrated to be accurate, precise, specific, and reproducible. While no matrix influence was detected, a list of interfering substances was compiled as a guideline for proper collection and storage of patient CSF samples. In addition, a set of recommendations on result interpretation is provided.
CONCLUSIONS:
This SMC assay is a robust and ultrasensitive method for the relative quantification of mHTT in human CSF. -
Polyglutamine expansion affects huntingtin conformation in multiple Huntington’s disease models
Scientific Reports
Conformational changes in disease-associated or mutant proteins represent a key pathological aspect of Huntington's disease (HD) and other protein misfolding diseases. Using immunoassays and biophysical approaches, we and others have recently reported that polyglutamine expansion in purified or recombinantly expressed huntingtin (HTT) proteins affects their conformational properties in a manner dependent on both polyglutamine repeat length and temperature but independent of HTT protein fragment…
Conformational changes in disease-associated or mutant proteins represent a key pathological aspect of Huntington's disease (HD) and other protein misfolding diseases. Using immunoassays and biophysical approaches, we and others have recently reported that polyglutamine expansion in purified or recombinantly expressed huntingtin (HTT) proteins affects their conformational properties in a manner dependent on both polyglutamine repeat length and temperature but independent of HTT protein fragment length. These findings are consistent with the HD mutation affecting structural aspects of the amino-terminal region of the protein, and support the concept that modulating mutant HTT conformation might provide novel therapeutic and diagnostic opportunities. We now report that the same conformational TR-FRET based immunoassay detects polyglutamine- and temperature-dependent changes on the endogenously expressed HTT protein in peripheral tissues and post-mortem HD brain tissue, as well as in tissues from HD animal models. We also find that these temperature- and polyglutamine-dependent conformational changes are sensitive to bona-fide phosphorylation on S13 and S16 within the N17 domain of HTT. These findings provide key clinical and preclinical relevance to the conformational immunoassay, and provide supportive evidence for its application in the development of therapeutics aimed at correcting the conformation of polyglutamine-expanded proteins as well as the pharmacodynamics readouts to monitor their efficacy in preclinical models and in HD patients.
-
Conformational modulation mediated by polyglutamine expansion in CAG repeat expansion disease-associated proteins.
Biochemical and Biophysical Research Communications
We have previously reported TR-FRET based immunoassays to detect a conformational change imparted on huntingtin protein by the polyglutamine expansion, which we confirmed using biophysical methodologies. Using these immunoassays, we now report that polyglutamine expansion influences the conformational properties of other polyglutamine disease proteins, exemplified by the androgen receptor (associated with spinal bulbar muscular atrophy) and TATA binding protein (associated with spinocerebellar…
We have previously reported TR-FRET based immunoassays to detect a conformational change imparted on huntingtin protein by the polyglutamine expansion, which we confirmed using biophysical methodologies. Using these immunoassays, we now report that polyglutamine expansion influences the conformational properties of other polyglutamine disease proteins, exemplified by the androgen receptor (associated with spinal bulbar muscular atrophy) and TATA binding protein (associated with spinocerebellar ataxia 17). Using artificial constructs bearing short or long polyglutamine expansions or a multimerized, unrelated epitope (mimicking the increase in anti-polyglutamine antibody epitopes present in polyglutamine repeats of increasing length) we confirmed that the conformational TR-FRET based immunoassay detects an intrinsic conformational property of polyglutamine repeats. The TR-FRET based conformational immunoassay may represent a rapid, scalable tool to identify modulators of polyglutamine-mediated conformational change in different proteins associated with CAG triplet repeat disorders.
-
Polyglutamine- and temperature-dependent conformational rigidity in mutant huntingtin revealed by immunoassays and circular dichroism spectroscopy.
PLoS One
In Huntington's disease, expansion of a CAG triplet repeat occurs in exon 1 of the huntingtin gene (HTT), resulting in a protein bearing>35 polyglutamine residues whose N-terminal fragments display a high propensity to misfold and aggregate. Recent data demonstrate that polyglutamine expansion results in conformational changes in the huntingtin protein (HTT), which likely influence its biological and biophysical properties. Developing assays to characterize and measure these conformational…
In Huntington's disease, expansion of a CAG triplet repeat occurs in exon 1 of the huntingtin gene (HTT), resulting in a protein bearing>35 polyglutamine residues whose N-terminal fragments display a high propensity to misfold and aggregate. Recent data demonstrate that polyglutamine expansion results in conformational changes in the huntingtin protein (HTT), which likely influence its biological and biophysical properties. Developing assays to characterize and measure these conformational changes in isolated proteins and biological samples would advance the testing of novel therapeutic approaches aimed at correcting mutant HTT misfolding. Time-resolved Förster energy transfer (TR-FRET)-based assays represent high-throughput, homogeneous, sensitive immunoassays widely employed for the quantification of proteins of interest. TR-FRET is extremely sensitive to small distances and can therefore provide conformational information based on detection of exposure and relative position of epitopes present on the target protein as recognized by selective antibodies. We have previously reported TR-FRET assays to quantify HTT proteins based on the use of antibodies specific for different amino-terminal HTT epitopes. Here, we investigate the possibility of interrogating HTT protein conformation using these assays.
Lingue
-
Italiano
Conoscenza madrelingua o bilingue
-
Inglese
Conoscenza professionale
Altre attività di Manuel
-
While part of our team is actively engaging with peers at EFMC-ISMC in Italy, our exceptional BD members, including Betsy McKibben and Michael…
While part of our team is actively engaging with peers at EFMC-ISMC in Italy, our exceptional BD members, including Betsy McKibben and Michael…
Consigliato da Manuel Daldin
-
A mix of excitement and pride when reading our own name in a #Nature journal article and so grateful to David Rubinsztein for giving me such an…
A mix of excitement and pride when reading our own name in a #Nature journal article and so grateful to David Rubinsztein for giving me such an…
Consigliato da Manuel Daldin
-
I have been working on #doubleblind #clinicaltrials on #neurodegenerativediseases for about 25 years and I am always surprised by what I observe once…
I have been working on #doubleblind #clinicaltrials on #neurodegenerativediseases for about 25 years and I am always surprised by what I observe once…
Consigliato da Manuel Daldin
-
Rob Haselberg is een bekend gezicht in Huntington-land. Hij ervaart de ziekte vanuit verschillende rollen: zoon, gendrager, mantelzorger, vader…
Rob Haselberg is een bekend gezicht in Huntington-land. Hij ervaart de ziekte vanuit verschillende rollen: zoon, gendrager, mantelzorger, vader…
Consigliato da Manuel Daldin
-
𝗛𝗶𝗴𝗵-𝗰𝗼𝗻𝘁𝗲𝗻𝘁 𝗰𝗲𝗹𝗹 𝗶𝗺𝗮𝗴𝗶𝗻𝗴 is crucial in drug discovery, enabling detailed analysis of cellular responses. Integrating…
𝗛𝗶𝗴𝗵-𝗰𝗼𝗻𝘁𝗲𝗻𝘁 𝗰𝗲𝗹𝗹 𝗶𝗺𝗮𝗴𝗶𝗻𝗴 is crucial in drug discovery, enabling detailed analysis of cellular responses. Integrating…
Consigliato da Manuel Daldin
-
Comunque dire: "Ho lasciato il posto fisso per iniziare nuove avventure" non vale se hai 10 case intestate in affitto, i tuoi genitori possiedono una…
Comunque dire: "Ho lasciato il posto fisso per iniziare nuove avventure" non vale se hai 10 case intestate in affitto, i tuoi genitori possiedono una…
Consigliato da Manuel Daldin
-
Really excited to share a preprint from our lab describing the genetic analysis of 9 HD GWAS/mismatch repair (MMR) genes in the Q140 knockin mouse…
Really excited to share a preprint from our lab describing the genetic analysis of 9 HD GWAS/mismatch repair (MMR) genes in the Q140 knockin mouse…
Consigliato da Manuel Daldin
-
See my thoughts on the recently reported results from the Uniqure AMT-130 Phase 1/2 clinical trial for the treatment of HD. https://lnkd.in/g3FjSbDe
See my thoughts on the recently reported results from the Uniqure AMT-130 Phase 1/2 clinical trial for the treatment of HD. https://lnkd.in/g3FjSbDe
Consigliato da Manuel Daldin
-
News from Uniqure’s AMT130 trial in HD.
News from Uniqure’s AMT130 trial in HD.
Consigliato da Manuel Daldin
-
According to ITA (the Italian Trade Agency), 𝗜𝘁𝗮𝗹𝘆 𝗵𝗮𝘀 𝘁𝗵𝗲 𝗯𝗲𝘀𝘁 𝗮𝗻𝗱 𝗳𝗮𝘀𝘁𝗲𝘀𝘁-𝗴𝗿𝗼𝘄𝗶𝗻𝗴 𝗟𝗶𝗳𝗲 𝗦𝗰𝗶𝗲𝗻𝗰𝗲…
According to ITA (the Italian Trade Agency), 𝗜𝘁𝗮𝗹𝘆 𝗵𝗮𝘀 𝘁𝗵𝗲 𝗯𝗲𝘀𝘁 𝗮𝗻𝗱 𝗳𝗮𝘀𝘁𝗲𝘀𝘁-𝗴𝗿𝗼𝘄𝗶𝗻𝗴 𝗟𝗶𝗳𝗲 𝗦𝗰𝗶𝗲𝗻𝗰𝗲…
Consigliato da Manuel Daldin
-
Do not hesitate to connect with Martina Bischetti also if you want to hear more about available #NMR positions in our group 😉 stay tuned!
Do not hesitate to connect with Martina Bischetti also if you want to hear more about available #NMR positions in our group 😉 stay tuned!
Consigliato da Manuel Daldin