Identification of protein quality control regulators using a Drosophila model of TPI deficiency

Hrizo, Stacy; Eicher, Samantha L; Myers, Tracey; McGrath, J.C.; Wodrich, Andrew P.K.; Venkatesh, Hemanth; Manjooran, Daniel T.; Swoger, Sabrina; Gagnon, Kim; Bruskin, Matthew; Lebedev, Maria V.; Zheng, Sherry Li; Vitantonio, Ana; Kim, Sung-Youn; Lamb, Zachary J.; Vogt, Andreas; Ruzhnikov, Maura R.Z.; Palladino, Michael J.

doi:10.1016/j.nbd.2021.105299

Cited by 5 publications

(9 citation statements)

References 41 publications

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“…Brilliant green, exhaustive diagnostic approach; Green, requiring other approaches to confirm the diagnosis; gray, insufficient laboratory markers to reach the diagnosis. (Garcia-Gomez et al, 2016;Grace et al, 2019;Dessy-Rodriguez et al, 2020;Hrizo et al, 2021).…”

Section: Discussionmentioning

confidence: 99%

Targeted Next Generation Sequencing and Diagnosis of Congenital Hemolytic Anemias: A Three Years Experience Monocentric Study

et al. 2021

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Congenital hemolytic anemias (CHAs) are heterogeneous and rare disorders caused by alterations in structure, membrane transport, metabolism, or red blood cell production. The pathophysiology of these diseases, in particular the rarest, is often poorly understood, and easy-to-apply tools for diagnosis, clinical management, and patient stratification are still lacking. We report the 3-years monocentric experience with a 43 genes targeted Next Generation Sequencing (t-NGS) panel in diagnosis of CHAs; 122 patients from 105 unrelated families were investigated and the results compared with conventional laboratory pathway. Patients were divided in two groups: 1) cases diagnosed with hematologic investigations to be confirmed at molecular level, and 2) patients with unexplained anemia after extensive hematologic investigation. The overall sensitivity of t-NGS was 74 and 35% for families of groups 1 and 2, respectively. Inside this cohort of patients we identified 26 new pathogenic variants confirmed by functional evidence. The implementation of laboratory work-up with t-NGS increased the number of diagnoses in cases with unexplained anemia; cytoskeleton defects are well detected by conventional tools, deserving t-NGS to atypical cases; the diagnosis of Gardos channelopathy, some enzyme deficiencies, familial siterosterolemia, X-linked defects in females and other rare and ultra-rare diseases definitely benefits of t-NGS approaches.

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Section: Discussionmentioning

confidence: 99%

Targeted Next Generation Sequencing and Diagnosis of Congenital Hemolytic Anemias: A Three Years Experience Monocentric Study

et al. 2021

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“…The structural basis for decreased protein stability is not known; however, a recent genetic screen for genes that regulate TPI stability has suggested that lower stability of mutant TPI is governed by a variety of proteins, including known PQC regulators, but also cotranslational modulators and a variety of proteins with unknown functions. 11 Therefore, there is no basis for developing targeted therapies. Thus, we reasoned that a phenotypic assay for TPI expression would be most appropriate to discover novel TPI Df therapeutics because such an assay would capture all mechanisms that lead to stabilization of mutant TPI.…”

Section: Discussionmentioning

confidence: 99%

“…We have recently used mutant TPI in a powerful Drosophila genetic screen to elucidate all (or at least most) of the PQC components that regulate TPI turnover. 11 We performed a genome-wide RNAi screen of all known and predicted components of the ubiquitin–proteasome system (UPS) and PQC pathways and numerous unknown proteins with domains similar to those commonly found in UPS and PQC components. Importantly, the screen identified ~27 proteins that are critical to TPI stability (including many that were not previously known to be involved in its turnover), novel proteins with no known or associated PQC function for any substrate, including some that could contribute to protein folding or ubiquitination, or are likely to be involved in co-translational PQC.…”

Section: Introductionmentioning

confidence: 99%

“…Importantly, the screen identified ~27 proteins that are critical to TPI stability (including many that were not previously known to be involved in its turnover), novel proteins with no known or associated PQC function for any substrate, including some that could contribute to protein folding or ubiquitination, or are likely to be involved in co-translational PQC. 11 The majority of these potential targets are known to be much less promiscuous than Hsp70 and Hsp90, which are known to modulate >75% of the proteome, suggesting that discovery of less toxic agents that stabilize mutant TPI should be feasible. 11…”

Section: Introductionmentioning

confidence: 99%

“…11 The majority of these potential targets are known to be much less promiscuous than Hsp70 and Hsp90, which are known to modulate >75% of the proteome, suggesting that discovery of less toxic agents that stabilize mutant TPI should be feasible. 11…”

Section: Introductionmentioning

confidence: 99%

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A High-Content Screening Assay for Small Molecules That Stabilize Mutant Triose Phosphate Isomerase (TPI) as Treatments for TPI Deficiency

et al. 2021

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Triose phosphate isomerase deficiency (TPI Df) is an untreatable, childhood-onset glycolytic enzymopathy. Patients typically present with frequent infections, anemia, and muscle weakness that quickly progresses with severe neuromusclar dysfunction requiring aided mobility and often respiratory support. Life expectancy after diagnosis is typically ~5 years. There are several described pathogenic mutations that encode functional proteins; however, these proteins, which include the protein resulting from the “common” TPIE105D mutation, are unstable due to active degradation by protein quality control (PQC) pathways. Previous work has shown that elevating mutant TPI levels by genetic or pharmacological intervention can ameliorate symptoms of TPI Df in fruit flies. To identify compounds that increase levels of mutant TPI, we have developed a human embryonic kidney (HEK) stable knock-in model expressing the common TPI Df protein fused with green fluorescent protein (HEK TPIE105D-GFP). To directly address the need for lead TPI Df therapeutics, these cells were developed into an optical drug discovery platform that was implemented for high-throughput screening (HTS) and validated in 3-day variability tests, meeting HTS standards. We initially used this assay to screen the 446-member National Institutes of Health (NIH) Clinical Collection and validated two of the hits in dose–response, by limited structure–activity relationship studies with a small number of analogs, and in an orthogonal, non-optical assay in patient fibroblasts. The data form the basis for a large-scale phenotypic screening effort to discover compounds that stabilize TPI as treatments for this devastating childhood disease.

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Murine model of triosephosphate isomerase deficiency with anemia and severe neuromuscular dysfunction

Myers

Ferguson

Gliniak

et al. 2022

Current Research in Neurobiology

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Identification of protein quality control regulators using a Drosophila model of TPI deficiency

Cited by 5 publications

References 41 publications

Targeted Next Generation Sequencing and Diagnosis of Congenital Hemolytic Anemias: A Three Years Experience Monocentric Study

Targeted Next Generation Sequencing and Diagnosis of Congenital Hemolytic Anemias: A Three Years Experience Monocentric Study

A High-Content Screening Assay for Small Molecules That Stabilize Mutant Triose Phosphate Isomerase (TPI) as Treatments for TPI Deficiency

Murine model of triosephosphate isomerase deficiency with anemia and severe neuromuscular dysfunction

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