Rin
Homma
Preventing Stabilization of Androgen Receptor Through Inhibition of Interaction with PSMD14
Abstract profile. Full document pending author claim.
Authors:
Rin Homma, Haniya Sperling, Jonathan Tsai
Date Created:
2025-01-01
Course Title:
Professor:
Not specified
About Paper:
Nuclear hormone receptors are transcription factors which activate individually to determine whether they interacted with PSMD14. upon interaction with small signaling molecules. The androgen Each residue was mutated by site-directed mutagenesis. Mutated receptor (AR) is a nuclear hormone receptor which contributes to AR was expressed in cells of the androgen sensitive LN-Cap a broad range of cell functions, including growth and proliferation.rostate cancer cell line. Levels of AR in cells with the mutated Cancer cells, particularly prostate cancer, are driven by activatedresidues were measured at baseline, and after titrated treatment AR signaling in order to grow and metastasize. AR protein is with AR agonist testosterone. Mutations V713A, D730A, and stabilized following androgen binding. We found through genetic V734A appeared to induce the greatest destabilization of AR as screens that the enzyme PSMD14 stabilizes AR and predicts a result of blocked interactions with PSMD14. Addition to and that specific residues mediate this interaction. We pinpointed modification of the mutation library is an ongoing project, with specific residues in the AR hydrophobic cleft and mutated them future direction to be taken in therapeutic development. Obtaining the Structure of the Staphylococcus aureus Lipoteichoic Acid Polymerase Wyatt Jensen, Forrest Hammel, Suzanne Walker Harvard College | Leverett House | Chemistry | 2027 Antimicrobial resistance (AMR) presents a significant obstacle LtaS using cryogenic electron microscopy (cryo-EM) and cutting- to improving global health outcomes, playing a role in nearly edgebiochemicaltechniquestooptimizeforthechallengingnature 5 million deaths in 2021. Identification of new targets and of this membrane protein. Namely, we engineer rigid LtaS- development of effective antibiotics are vital steps in overcoming BRIL fusions that enable us to increase the apparent size of such a challenge. Methicillin-resistant Staphylococcus aureus LtaS using fiducial marks, which enhance our ability to obtain (MRSA) remains a high-priority bacterial pathogen due to its high a high-resolution structure by cryo-EM. We then embed these incidence and transmissibility, especially in clinical settings. S.fusions in lipid nanodiscs that mimic the native lipid environment aureus maintains a thick peptidoglycan cell wall with polymers of the S. aureus cell envelope. We demonstrate that LtaS can known as teichoic acids interwoven throughout, either covalently be successfully reconstituted in lipid nanodiscs at the milligram linked to the peptidoglycan as wall teichoic acids (WTAs) or to thescale, which is suitable for cryo-EM sample preparation and data outer leaflet of the lipid bilayer as lipoteichoic acids (LTAs). LTprocessing. Together, these approaches position us to soon resolve biosynthesis is essential for S. aureus growth and cell division, aa structure of LtaS that informs the design of LTA biosynthetic theenzymeresponsibleforLTAsynthesis, LtaS,isapromisingbut inhibitors and develops our understanding of LTA biosynthesis for elusive drug target. We therefore seek to obtain the structure of many Gram-positive bacterial pathogens. 102 Program for Research in Science and Engineering Microbial Disruption of Nitrergic Neurons in Gastrointestinal Motility Disorders Mira Jiang, Ariel Robinson, Meenakshi Rao Harvard College | Leverett House | Neuroscience | 2026 Many gastrointestinal (GI) motility disorders are characterized by at any timepoint between VEH and ABX mice (p = 0.9206), lossofinhibitorymotorneuronsinthemyentericplexus,markedby indicating that microbial depletion causes NOS1 downregulation, expression of the enzyme NOS1. In mouse models, administration not cell loss. Enteric neurons broadly segregate into nitrergic of broad-spectrum antibiotics slows GI transit and causes a and cholinergic classes, that are considered mutually exclusive. reduction in NOS1-immunoreactive neurons that is reversible with Calretinin marks a large subset of cholinergic neurons, including bacterial recolonization, suggesting gut microbes are essential forexcitatory motor neurons. ABX induced nearly a 3-fold increase the maintenance and regeneration of these neurons. To determine in the proportion of tdTomato+ neurons that expressed Calretinin underlying mechanisms, we genetically labeled NOS1 neurons in (29:9% ▯ 4:9%, n = 4) at 21 days compared to VEH controls adult Nos1CreER/+Rosa26 TdTomato/mice, exposed them to a broad- (10% ▯ 4:9%, n = 3). Similarly, the percentage of NOS1 + spectrum antibiotic cocktail (ABX) or vehicle (VEH) in drinking neurons that colocalized with Calretinin was increased in mice on water for 7-21 days, and then isolated colonic tissues for NOS1 both 7 days (26:3% ▯ 2%, n = 2) and 21 days of ABX (26:2% ▯ immunohistochemistry and confocal imaging. At 7 days, only 7:2%, n = 4) compared to VEH controls (13:4%▯6:9%, n = 3). 45.1% ± 10.5% of tdTomato myenteric neurons expressed NOS1 This dysregulation may explain ABX associated dysmotility and in ABX mice (n = 14), compared to 91:5% ▯ 5:4% in VEH suggests that NOS1 neurons are not readily lost with microbial controls (n = 8). This deficit was partially restored by 21 depletion, providing an opportunity to identify microbial signals days (81:1% ▯ 3%, n = 5) despite continued ABX exposure. that could be utilized to restore NOS1 expression in GI motility The proportion of tdTomato neurons, however, was no different disorders. Investigating the Influence of Geldanamycin on Temperature Preference in Drosophila melanogaster Kirsti Jones, Shraddha Lall, Benjamin de Bivort Emmanuel College, University of Cambridge | Veterinary Medicine | 2028 Individual fruit flies display variable preference even within the from previous literature. Temperature preference was measured samegenotype,yettheunderlyingmechanismsforthisbehavioural in custom thermal rigs where individual flies were presented with variability remain relatively unknown. Previous data on the 20 C and 28 C environments in each half of a corridor-like maze. chaperone protein spaghetti(spag) mutants suggest an increased Movement was tracked by a camera over 2 hours. We expect variability in temperature preference within populations, but temperature preference variability to be greater in GA-treated flies complementary pharmacological methods are yet to be tested. than controls in line with data obtained from previously mentioned Inhibition of chaperone proteins, which help other proteins fold, spag mutants. is hypothesised to ’unmask’ variability in protein structure, with Future work looks to consider further genetic contexts, possible functional implications. This project thus aims to including spag misexpression and RNA interference mutants to assess the impact of geldanamycin (GA), an Hsp90 inhibitor compare to current data. Temperature preference as an example that interferes with spag function, on behavioural variability in Drosophila melanogaster. of behavioural variability may provide insights into underlying mechanisms of personality determination of individuals within Flies were raised on food containing either 1.5 ▯g/mL GA or populations. dimethyl sulfoxide solvent, where GA dosage was determined Investigating the Role of TXNIP and ER Stress in Modulating P53-Mediated ChangesinHumanInducedPluripotentStemCell-DerivedCardiacOrganoids Wan Ting Ke, Nivedhitha Velayutham, Richard Lee Harvard College | Currier House | Human Developmental and Regenerative Biology | 2028 Cardiovascular diseases are the leading cause of death in modulating p53-mediated changes in hiPSC-derived cardiac the United States. Unlike many other cell types, adult organoids. For my summer project, three-dimensional cultures cardiomyocytes cannot regenerate as they have exited the cell of hiPSC-CMs were treated with Nutlin-3a (p53 activator), SRI- cycle. Instead, damaged cardiomyocytes are replaced with fibrotic 37330 (TXNIP inhibitor), and 4 Phenylbutyric acid (ER stress scar tissue, permanently decreasing the contractile ability of the inhibitor), alone or in-combination, starting from day 9 of heart. Current efforts aim at replenishing lost or damaged differentiation. Reverse transcription-quantitative PCR was then cardiomyocytes by transplanting human induced pluripotent stem performed to compare the expression of key marker genes such as cell-derived cardiomyocytes (hiPSC-CMs). However, hiPSC- TP53, TXNIP, CALR, TNNT2, and DDIT3 to elucidate whether CMs exhibit “fetal-like” phenotypic characteristics, leading SRI-37330 and 4PBA modulate p53, TXNIP and ER stress. to ventricular arrhythmias when engrafted in animal models. While scRNAseq data suggest that TXNIP and ER stress lie Previously, the lab demonstrated that p53 activation promoted downstream of p53, preliminary results are inconclusive regarding cardiomyocytematurationbyFOXO-FOXM1regulationinhiPSC- the effects of inhibiting TXNIP and 4PBA. For instance, the use derived cardiac organoids. Ongoing work based on single of SRI-37330 either significantly upregulates or downregulates the cell RNA sequencing (scRNAseq) of Nutlin-3a treated cardiac gene expressions of TXNIP in different cell preparation batches, organoids show that thioredoxin-interacting protein (TXNIP) suggesting that these inhibitors may not be working on an RNA and endoplasmic reticulum (ER) stress markers are significantly level. Thus, it may be necessary to perform protein assessment upregulated in p53-activated groups. This study seeks to via Western blotting and immunostaining to accurately identify the understand the individual roles of TXNIP and ER Stress in impact of TXNIP or ER stress inhibition on hiPSC-CMs.
Abstract:
Nuclear hormone receptors are transcription factors which activate individually to determine whether they interacted with PSMD14. upon interaction with small signaling molecules. The androgen Each residue was mutated by site-directed mutagenesis. Mutated receptor (AR) is a nuclear hormone receptor which contributes to AR was expressed in cells of the androgen sensitive LN-Cap a broad range of cell functions, including growth and proliferation.rostate cancer cell line. Levels of AR in cells with the mutated Cancer cells, particularly prostate cancer, are driven by activatedresidues were measured at baseline, and after titrated treatment AR signaling in order to grow and metastasize. AR protein is with AR agonist testosterone. Mutations V713A, D730A, and stabilized following androgen binding. We found through genetic V734A appeared to induce the greatest destabilization of AR as screens that the enzyme PSMD14 stabilizes AR and predicts a result of blocked interactions with PSMD14. Addition to and that specific residues mediate this interaction. We pinpointed modification of the mutation library is an ongoing project, with specific residues in the AR hydrophobic cleft and mutated them future direction to be taken in therapeutic development. Obtaining the Structure of the Staphylococcus aureus Lipoteichoic Acid Polymerase Wyatt Jensen, Forrest Hammel, Suzanne Walker Harvard College | Leverett House | Chemistry | 2027 Antimicrobial resistance (AMR) presents a significant obstacle LtaS using cryogenic electron microscopy (cryo-EM) and cutting- to improving global health outcomes, playing a role in nearly edgebiochemicaltechniquestooptimizeforthechallengingnature 5 million deaths in 2021. Identification of new targets and of this membrane protein. Namely, we engineer rigid LtaS- development of effective antibiotics are vital steps in overcoming BRIL fusions that enable us to increase the apparent size of such a challenge. Methicillin-resistant Staphylococcus aureus LtaS using fiducial marks, which enhance our ability to obtain (MRSA) remains a high-priority bacterial pathogen due to its high a high-resolution structure by cryo-EM. We then embed these incidence and transmissibility, especially in clinical settings. S.fusions in lipid nanodiscs that mimic the native lipid environment aureus maintains a thick peptidoglycan cell wall with polymers of the S. aureus cell envelope. We demonstrate that LtaS can known as teichoic acids interwoven throughout, either covalently be successfully reconstituted in lipid nanodiscs at the milligram linked to the peptidoglycan as wall teichoic acids (WTAs) or to thescale, which is suitable for cryo-EM sample preparation and data outer leaflet of the lipid bilayer as lipoteichoic acids (LTAs). LTprocessing. Together, these approaches position us to soon resolve biosynthesis is essential for S. aureus growth and cell division, aa structure of LtaS that informs the design of LTA biosynthetic theenzymeresponsibleforLTAsynthesis, LtaS,isapromisingbut inhibitors and develops our understanding of LTA biosynthesis for elusive drug target. We therefore seek to obtain the structure of many Gram-positive bacterial pathogens. 102 Program for Research in Science and Engineering Microbial Disruption of Nitrergic Neurons in Gastrointestinal Motility Disorders Mira Jiang, Ariel Robinson, Meenakshi Rao Harvard College | Leverett House | Neuroscience | 2026 Many gastrointestinal (GI) motility disorders are characterized by at any timepoint between VEH and ABX mice (p = 0.9206), lossofinhibitorymotorneuronsinthemyentericplexus,markedby indicating that microbial depletion causes NOS1 downregulation, expression of the enzyme NOS1. In mouse models, administration not cell loss. Enteric neurons broadly segregate into nitrergic of broad-spectrum antibiotics slows GI transit and causes a and cholinergic classes, that are considered mutually exclusive. reduction in NOS1-immunoreactive neurons that is reversible with Calretinin marks a large subset of cholinergic neurons, including bacterial recolonization, suggesting gut microbes are essential forexcitatory motor neurons. ABX induced nearly a 3-fold increase the maintenance and regeneration of these neurons. To determine in the proportion of tdTomato+ neurons that expressed Calretinin underlying mechanisms, we genetically labeled NOS1 neurons in (29:9% ▯ 4:9%, n = 4) at 21 days compared to VEH controls adult Nos1CreER/+Rosa26 TdTomato/mice, exposed them to a broad- (10% ▯ 4:9%, n = 3). Similarly, the percentage of NOS1 + spectrum antibiotic cocktail (ABX) or vehicle (VEH) in drinking neurons that colocalized with Calretinin was increased in mice on water for 7-21 days, and then isolated colonic tissues for NOS1 both 7 days (26:3% ▯ 2%, n = 2) and 21 days of ABX (26:2% ▯ immunohistochemistry and confocal imaging. At 7 days, only 7:2%, n = 4) compared to VEH controls (13:4%▯6:9%, n = 3). 45.1% ± 10.5% of tdTomato myenteric neurons expressed NOS1 This dysregulation may explain ABX associated dysmotility and in ABX mice (n = 14), compared to 91:5% ▯ 5:4% in VEH suggests that NOS1 neurons are not readily lost with microbial controls (n = 8). This deficit was partially restored by 21 depletion, providing an opportunity to identify microbial signals days (81:1% ▯ 3%, n = 5) despite continued ABX exposure. that could be utilized to restore NOS1 expression in GI motility The proportion of tdTomato neurons, however, was no different disorders. Investigating the Influence of Geldanamycin on Temperature Preference in Drosophila melanogaster Kirsti Jones, Shraddha Lall, Benjamin de Bivort Emmanuel College, University of Cambridge | Veterinary Medicine | 2028 Individual fruit flies display variable preference even within the from previous literature. Temperature preference was measured samegenotype,yettheunderlyingmechanismsforthisbehavioural in custom thermal rigs where individual flies were presented with variability remain relatively unknown. Previous data on the 20 C and 28 C environments in each half of a corridor-like maze. chaperone protein spaghetti(spag) mutants suggest an increased Movement was tracked by a camera over 2 hours. We expect variability in temperature preference within populations, but temperature preference variability to be greater in GA-treated flies complementary pharmacological methods are yet to be tested. than controls in line with data obtained from previously mentioned Inhibition of chaperone proteins, which help other proteins fold, spag mutants. is hypothesised to ’unmask’ variability in protein structure, with Future work looks to consider further genetic contexts, possible functional implications. This project thus aims to including spag misexpression and RNA interference mutants to assess the impact of geldanamycin (GA), an Hsp90 inhibitor compare to current data. Temperature preference as an example that interferes with spag function, on behavioural variability in Drosophila melanogaster. of behavioural variability may provide insights into underlying mechanisms of personality determination of individuals within Flies were raised on food containing either 1.5 ▯g/mL GA or populations. dimethyl sulfoxide solvent, where GA dosage was determined Investigating the Role of TXNIP and ER Stress in Modulating P53-Mediated ChangesinHumanInducedPluripotentStemCell-DerivedCardiacOrganoids Wan Ting Ke, Nivedhitha Velayutham, Richard Lee Harvard College | Currier House | Human Developmental and Regenerative Biology | 2028 Cardiovascular diseases are the leading cause of death in modulating p53-mediated changes in hiPSC-derived cardiac the United States. Unlike many other cell types, adult organoids. For my summer project, three-dimensional cultures cardiomyocytes cannot regenerate as they have exited the cell of hiPSC-CMs were treated with Nutlin-3a (p53 activator), SRI- cycle. Instead, damaged cardiomyocytes are replaced with fibrotic 37330 (TXNIP inhibitor), and 4 Phenylbutyric acid (ER stress scar tissue, permanently decreasing the contractile ability of the inhibitor), alone or in-combination, starting from day 9 of heart. Current efforts aim at replenishing lost or damaged differentiation. Reverse transcription-quantitative PCR was then cardiomyocytes by transplanting human induced pluripotent stem performed to compare the expression of key marker genes such as cell-derived cardiomyocytes (hiPSC-CMs). However, hiPSC- TP53, TXNIP, CALR, TNNT2, and DDIT3 to elucidate whether CMs exhibit “fetal-like” phenotypic characteristics, leading SRI-37330 and 4PBA modulate p53, TXNIP and ER stress. to ventricular arrhythmias when engrafted in animal models. While scRNAseq data suggest that TXNIP and ER stress lie Previously, the lab demonstrated that p53 activation promoted downstream of p53, preliminary results are inconclusive regarding cardiomyocytematurationbyFOXO-FOXM1regulationinhiPSC- the effects of inhibiting TXNIP and 4PBA. For instance, the use derived cardiac organoids. Ongoing work based on single of SRI-37330 either significantly upregulates or downregulates the cell RNA sequencing (scRNAseq) of Nutlin-3a treated cardiac gene expressions of TXNIP in different cell preparation batches, organoids show that thioredoxin-interacting protein (TXNIP) suggesting that these inhibitors may not be working on an RNA and endoplasmic reticulum (ER) stress markers are significantly level. Thus, it may be necessary to perform protein assessment upregulated in p53-activated groups. This study seeks to via Western blotting and immunostaining to accurately identify the understand the individual roles of TXNIP and ER Stress in impact of TXNIP or ER stress inhibition on hiPSC-CMs.
Source:
Harvard / Harvard College | Quincy House | Chemistry | 2027 / 2025
Topics:
cell, neuron, txnip, nos1, protein, ltas, abx, stres, p53, days, preference, variability
