A5024695166- Protein Degradation and Inhibitors
- Histone Deacetylase Inhibitors Research
- Acute Myeloid Leukemia Research
- Cancer-related gene regulation
- Myeloproliferative Neoplasms: Diagnosis and Treatment
- Epigenetics and DNA Methylation
- Cancer Genomics and Diagnostics
- Computational Drug Discovery Methods
- CAR-T cell therapy research
- Chronic Myeloid Leukemia Treatments
- Immune Cell Function and Interaction
- Immune cells in cancer
- Single-cell and spatial transcriptomics
- Statistical Methods in Clinical Trials
- Multiple Myeloma Research and Treatments
- CRISPR and Genetic Engineering
- Hemoglobinopathies and Related Disorders
Vanderbilt University
2024
Vanderbilt University Medical Center
2022-2024
Pennsylvania State University
2023
Clonal hematopoiesis (CH) is an age-associated phenomenon that increases the risk of hematologic malignancy and cardiovascular disease. CH thought to enhance disease through inflammation in peripheral blood.1 Here, we profile blood gene expression 66 968 single cells from a cohort 17 patients with 7 controls. Using novel mitochondrial DNA barcoding approach, were able identify separately compare mutant Tet methylcytosine dioxygenase 2 (TET2) methyltransferase 3A (DNMT3A) nonmutant...
Abstract Impairing the BET family coactivator BRD4 with small-molecule inhibitors (BETi) showed encouraging preclinical activity in treating acute myeloid leukemia (AML). However, dose-limiting toxicities and limited clinical dampened enthusiasm for BETi as a single agent. resistance AML myeloblasts was found to correlate maintaining mitochondrial respiration, suggesting that identifying metabolic pathway sustaining integrity could help develop approaches improve efficacy. Herein, we...
Clonal hematopoiesis (CH) is an age-associated phenomenon leading to increased risk of both hematologic malignancy and nonmalignant organ dysfunction. Increasingly available genetic testing has made the incidental discovery CH clinically common yet evidence-based guidelines effective management strategies prevent adverse health outcomes are lacking. To address this gap, prospective CHIVE (clonal inflammation in vasculature) registry biorepository was created identify monitor individuals at...
Abstract Clonal hematopoiesis (CH) is an age-associated phenomenon that increases risk for hematologic malignancy and cardiovascular disease. CH thought to enhance disease through inflammation in the peripheral blood 1 . Here, we profile gene expression 66,968 single cells from a cohort of 17 patients 7 controls. Using novel mitochondrial DNA barcoding approach, were able identify separately compare mutant TET2 DNMT3A non-mutant counterparts. We discovered vast majority mutated myeloid...
<p>Table S1: Metabolomics Raw Data</p>
<p>Figure S5: Oxamate displays minimal toxicity to AML myeloblasts alone. were cultured for 72 hr with BETi (0.15 µM) and indicated lactate utilization inhibitors (AZD3965, UK5099, oxamate = 0.1 viability quantified using a fluorescent plate reader. Viability was relative cells treated vehicle control. Each point represents the mean (technical duplicate) value obtained from an individual experiment (n=3).</p>
<p>Figure S1: Glutaminolysis and fatty acid oxidation does not allow MOLM-13 to metabolically bypass BET inhibition. (A) MV-4-11 cells were cultured for 72 hr with BETi (0.15 µM) viability quantified using a fluorescent plate reader. Viability was relative treated vehicle control. Each point represents the mean (technical triplicate) value obtained from an individual experiment (n=3). (B-E) loaded CellTrace dye either (B-C) V-0302 (25 or (D-E) etomoxir (150 µM). (B, D) To assess...
<p>Table S2: Characteristics of AML patient samples tested with BETi.</p>
<p>Table S1: Metabolomics Raw Data</p>
<div>Abstract<p>Impairing the BET family coactivator BRD4 with small-molecule inhibitors (BETi) showed encouraging preclinical activity in treating acute myeloid leukemia (AML). However, dose-limiting toxicities and limited clinical dampened enthusiasm for BETi as a single agent. resistance AML myeloblasts was found to correlate maintaining mitochondrial respiration, suggesting that identifying metabolic pathway sustaining integrity could help develop approaches improve efficacy....
<p>Figure S3: CRISPR technology disrupts target genes. Expressions of (A) BRD4, (B, C) LDHB, and (C) MCT1 were disrupted in the indicated cell lines by CRISPR. The percent indel efficiency genetic disruption was quantified TIDE analysis Sanger sequencing relative to scramble control after 72 hr. (B) Cells treated with BETi (0.15 uM) for 48 hr viability a fluorescence plate reader. Each point represents mean value obtained from an individual experiment (n=3). (D) transfected siRNA...
<p>Figure S3: CRISPR technology disrupts target genes. Expressions of (A) BRD4, (B, C) LDHB, and (C) MCT1 were disrupted in the indicated cell lines by CRISPR. The percent indel efficiency genetic disruption was quantified TIDE analysis Sanger sequencing relative to scramble control after 72 hr. (B) Cells treated with BETi (0.15 uM) for 48 hr viability a fluorescence plate reader. Each point represents mean value obtained from an individual experiment (n=3). (D) transfected siRNA...
<p>Figure S4: UK5099 does not enhance the efficacy of BETi in preventing MOLM-13 chimerism. NSGS mice were engrafted with cells and treated vehicle control, (50 mg/kg) 5 times a week, twice daily and/or (40 three once daily. After 2 wks, peripheral blood (BL), spleen (SPL), bone marrow (BM) harvested for chimerism analysis. Each point represents single mouse (n = 5). Two-way ANOVA Tukey multiple comparisons test (*P ≤ 0.05, **P 0.01, ***P 0.001, ****P 0.0001, ns significant).</p>
<p>Figure S2: Maximal respiration in MOLM-13 cells is unaffected by the addition of BETi and/or lactate utilization inhibitors. MV-4-11 and were cultured for 72 hr with indicated treatments (BETi = 0.15 µM; AZD3965, UK5099, oxamate 0.1 µM), subjected to a mitochondrial stress test, oxygen consumption rates (OCR) quantified extracellular flux analysis. was where each point represents individual experiments (n=3). Two-way ANOVA Sidak’s multiple comparisons test (*P ≤ 0.05, ns not...
<p>Table S2: Characteristics of AML patient samples tested with BETi.</p>
<p>Figure S2: Maximal respiration in MOLM-13 cells is unaffected by the addition of BETi and/or lactate utilization inhibitors. MV-4-11 and were cultured for 72 hr with indicated treatments (BETi = 0.15 µM; AZD3965, UK5099, oxamate 0.1 µM), subjected to a mitochondrial stress test, oxygen consumption rates (OCR) quantified extracellular flux analysis. was where each point represents individual experiments (n=3). Two-way ANOVA Sidak’s multiple comparisons test (*P ≤ 0.05, ns not...
<div>Abstract<p>Impairing the BET family coactivator BRD4 with small-molecule inhibitors (BETi) showed encouraging preclinical activity in treating acute myeloid leukemia (AML). However, dose-limiting toxicities and limited clinical dampened enthusiasm for BETi as a single agent. resistance AML myeloblasts was found to correlate maintaining mitochondrial respiration, suggesting that identifying metabolic pathway sustaining integrity could help develop approaches improve efficacy....
<p>Figure S5: Oxamate displays minimal toxicity to AML myeloblasts alone. were cultured for 72 hr with BETi (0.15 µM) and indicated lactate utilization inhibitors (AZD3965, UK5099, oxamate = 0.1 viability quantified using a fluorescent plate reader. Viability was relative cells treated vehicle control. Each point represents the mean (technical duplicate) value obtained from an individual experiment (n=3).</p>
<p>Figure S4: UK5099 does not enhance the efficacy of BETi in preventing MOLM-13 chimerism. NSGS mice were engrafted with cells and treated vehicle control, (50 mg/kg) 5 times a week, twice daily and/or (40 three once daily. After 2 wks, peripheral blood (BL), spleen (SPL), bone marrow (BM) harvested for chimerism analysis. Each point represents single mouse (n = 5). Two-way ANOVA Tukey multiple comparisons test (*P ≤ 0.05, **P 0.01, ***P 0.001, ****P 0.0001, ns significant).</p>
<p>Figure S1: Glutaminolysis and fatty acid oxidation does not allow MOLM-13 to metabolically bypass BET inhibition. (A) MV-4-11 cells were cultured for 72 hr with BETi (0.15 µM) viability quantified using a fluorescent plate reader. Viability was relative treated vehicle control. Each point represents the mean (technical triplicate) value obtained from an individual experiment (n=3). (B-E) loaded CellTrace dye either (B-C) V-0302 (25 or (D-E) etomoxir (150 µM). (B, D) To assess...
CRISPR-Cas9 is a useful tool for inserting precise genetic alterations through homology-directed repair (HDR), although current methods rely on provision of an exogenous template. Here, we tested the possibility repairing heterozygous single nucleotide variants (SNVs) using cell′s own wild-type allele rather than Using high-fidelity Cas9 to perform allele-specific CRISPR across multiple human leukemia cell lines as well in primary hematopoietic cells from patients with leukemia, find high...
Drug repositioning seeks to leverage existing clinical knowledge identify alternative settings for approved drugs. However, efforts fail demonstrate improved success rates in late-stage trials. Focusing on 11 kinase inhibitors that have been evaluated 139 hypotheses, we use data mining characterize the state of repurposing. Then, using a simple experimental correction with human serum proteins vitro pharmacodynamic assays, develop measurement drug's effective exposure. We show this metric is...