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Article ; Online: Localization of PPM1H phosphatase tunes Parkinson's disease-linked LRRK2 kinase-mediated Rab GTPase phosphorylation and ciliogenesis.

Yeshaw, Wondwossen M / Adhikari, Ayan / Chiang, Claire Y / Dhekne, Herschel S / Wawro, Paulina S / Pfeffer, Suzanne R

Proceedings of the National Academy of Sciences of the United States of America

2023 Volume 120, Issue 44, Page(s) e2315171120

Abstract: PPM1H phosphatase reverses Parkinson's disease-associated, Leucine Rich Repeat Kinase 2-mediated Rab GTPase phosphorylation. We show here that PPM1H relies on an N-terminal amphipathic helix for Golgi localization. The amphipathic helix enables PPM1H to ... ...

Abstract	PPM1H phosphatase reverses Parkinson's disease-associated, Leucine Rich Repeat Kinase 2-mediated Rab GTPase phosphorylation. We show here that PPM1H relies on an N-terminal amphipathic helix for Golgi localization. The amphipathic helix enables PPM1H to bind to liposomes in vitro, and small, highly curved liposomes stimulate PPM1H activity. We artificially anchored PPM1H to the Golgi, mitochondria, or mother centriole. Our data show that regulation of Rab10 GTPase phosphorylation requires PPM1H access to Rab10 at or near the mother centriole. Moreover, poor colocalization of Rab12 explains in part why it is a poor substrate for PPM1H in cells but not in vitro. These data support a model in which localization drives PPM1H substrate selection and centriolar PPM1H is critical for regulation of Rab GTPase-regulated ciliogenesis. Moreover, Golgi localized PPM1H may maintain active Rab GTPases on the Golgi to carry out their nonciliogenesis-related functions in membrane trafficking.
MeSH term(s)	Humans ; Phosphorylation ; Phosphoric Monoester Hydrolases/metabolism ; Parkinson Disease/genetics ; Parkinson Disease/metabolism ; Liposomes ; rab GTP-Binding Proteins/genetics ; rab GTP-Binding Proteins/metabolism ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/genetics ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/metabolism ; Phosphoprotein Phosphatases/metabolism
Chemical Substances	Phosphoric Monoester Hydrolases (EC 3.1.3.2) ; Liposomes ; rab GTP-Binding Proteins (EC 3.6.5.2) ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 (EC 2.7.11.1) ; LRRK2 protein, human (EC 2.7.11.1) ; PPM1H protein, human (EC 3.1.3.16) ; Phosphoprotein Phosphatases (EC 3.1.3.16)
Language	English
Publishing date	2023-10-27
Publishing country	United States
Document type	Journal Article
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.2315171120
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Article: Target gene regulatory network of miR-497 in angiosarcoma.

Benton, Annaleigh / Terwilliger, Emma / Moriarty, Noah M / Liu, Bozhi / Murphy, Ant / Maluvac, Hannah / Shu, Mae / Gartenhaus, Lauren E / Janson, Nimod D / Pfeffer, Claire M / Utturkar, Sagar M / Parkinson, Elizabeth I / Lanman, Nadia A / Hanna, Jason A

bioRxiv : the preprint server for biology

2023

Abstract: Angiosarcoma (AS) is a vascular sarcoma that is highly aggressive and metastatic. Due to its rarity, treatment options for patients are limited, therefore more research is needed to identify possible therapeutic vulnerabilities. We previously found that ... ...

Abstract	Angiosarcoma (AS) is a vascular sarcoma that is highly aggressive and metastatic. Due to its rarity, treatment options for patients are limited, therefore more research is needed to identify possible therapeutic vulnerabilities. We previously found that conditional deletion of
Language	English
Publishing date	2023-09-25
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.09.24.559218
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Article ; Online: Genome-wide screen reveals Rab12 GTPase as a critical activator of Parkinson's disease-linked LRRK2 kinase.

Dhekne, Herschel S / Tonelli, Francesca / Yeshaw, Wondwossen M / Chiang, Claire Y / Limouse, Charles / Jaimon, Ebsy / Purlyte, Elena / Alessi, Dario R / Pfeffer, Suzanne R

eLife

2023 Volume 12

Abstract: Activating mutations in the leucine-rich repeat kinase 2 (LRRK2) cause Parkinson's disease. LRRK2 phosphorylates a subset of Rab GTPases, particularly Rab10 and Rab8A, and we showed previously that these phosphoRabs play an important role in LRRK2 ... ...

Abstract	Activating mutations in the leucine-rich repeat kinase 2 (LRRK2) cause Parkinson's disease. LRRK2 phosphorylates a subset of Rab GTPases, particularly Rab10 and Rab8A, and we showed previously that these phosphoRabs play an important role in LRRK2 membrane recruitment and activation (Vides et al., 2022). To learn more about LRRK2 pathway regulation, we carried out an unbiased, CRISPR-based genome-wide screen to identify modifiers of cellular phosphoRab10 levels. A flow cytometry assay was developed to detect changes in phosphoRab10 levels in pools of mouse NIH-3T3 cells harboring unique CRISPR guide sequences. Multiple negative and positive regulators were identified; surprisingly, knockout of the
MeSH term(s)	Animals ; Mice ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/genetics ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/metabolism ; Mutation ; Parkinson Disease/genetics ; Parkinson Disease/metabolism ; Phosphorylation ; rab GTP-Binding Proteins/genetics ; rab GTP-Binding Proteins/metabolism
Chemical Substances	Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 (EC 2.7.11.1) ; rab GTP-Binding Proteins (EC 3.6.5.2) ; rab12 protein, mouse (EC 3.6.1.-)
Language	English
Publishing date	2023-10-24
Publishing country	England
Document type	Journal Article
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.87098
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Article ; Online: Nanoactuator for Neuronal Optoporation.

ACS nano

2024

Abstract: Light-driven modulation of neuronal activity at high spatial-temporal resolution is becoming of high interest in neuroscience. In addition to optogenetics, nongenetic membrane-targeted nanomachines that alter the electrical state of the neuronal ... ...

Abstract	Light-driven modulation of neuronal activity at high spatial-temporal resolution is becoming of high interest in neuroscience. In addition to optogenetics, nongenetic membrane-targeted nanomachines that alter the electrical state of the neuronal membranes are in demand. Here, we engineered and characterized a photoswitchable conjugated compound (BV-1) that spontaneously partitions into the neuronal membrane and undergoes a charge transfer upon light stimulation. The activity of primary neurons is not affected in the dark, whereas millisecond light pulses of cyan light induce a progressive decrease in membrane resistance and an increase in inward current matched to a progressive depolarization and action potential firing. We found that illumination of BV-1 induces oxidation of membrane phospholipids, which is necessary for the electrophysiological effects and is associated with decreased membrane tension and increased membrane fluidity. Time-resolved atomic force microscopy and molecular dynamics simulations performed on planar lipid bilayers revealed that the underlying mechanism is a light-driven formation of pore-like structures across the plasma membrane. Such a phenomenon decreases membrane resistance and increases permeability to monovalent cations, namely, Na
Language	English
Publishing date	2024-04-30
Publishing country	United States
Document type	Journal Article
ISSN	1936-086X
ISSN (online)	1936-086X
DOI	10.1021/acsnano.4c01672
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Article ; Online: Apoptosis: A Target for Anticancer Therapy.

Pfeffer, Claire M / Singh, Amareshwar T K

International journal of molecular sciences

2018 Volume 19, Issue 2

Abstract: Apoptosis, the cell's natural mechanism for death, is a promising target for anticancer therapy. Both the intrinsic and extrinsic pathways use caspases to carry out apoptosis through the cleavage of hundreds of proteins. In cancer, the apoptotic pathway ... ...

Abstract	Apoptosis, the cell's natural mechanism for death, is a promising target for anticancer therapy. Both the intrinsic and extrinsic pathways use caspases to carry out apoptosis through the cleavage of hundreds of proteins. In cancer, the apoptotic pathway is typically inhibited through a wide variety of means including overexpression of antiapoptotic proteins and under-expression of proapoptotic proteins. Many of these changes cause intrinsic resistance to the most common anticancer therapy, chemotherapy. Promising new anticancer therapies are plant-derived compounds that exhibit anticancer activity through activating the apoptotic pathway.
MeSH term(s)	Animals ; Antineoplastic Agents, Phytogenic/pharmacology ; Apoptosis/drug effects ; Carcinogenesis/drug effects ; Carcinogenesis/genetics ; Carcinogenesis/metabolism ; Carcinogenesis/pathology ; Caspases/genetics ; Caspases/metabolism ; Cell Line, Tumor ; Curcumin/pharmacology ; Fas Ligand Protein/genetics ; Fas Ligand Protein/metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Mice ; Molecular Targeted Therapy ; Neoplasms/drug therapy ; Neoplasms/genetics ; Neoplasms/metabolism ; Neoplasms/pathology ; Proto-Oncogene Proteins c-bcl-2/genetics ; Proto-Oncogene Proteins c-bcl-2/metabolism ; Signal Transduction ; bcl-2-Associated X Protein/genetics ; bcl-2-Associated X Protein/metabolism ; fas Receptor/genetics ; fas Receptor/metabolism
Chemical Substances	Antineoplastic Agents, Phytogenic ; BAX protein, human ; BCL2 protein, human ; FAS protein, human ; FASLG protein, human ; Fas Ligand Protein ; Proto-Oncogene Proteins c-bcl-2 ; bcl-2-Associated X Protein ; fas Receptor ; Caspases (EC 3.4.22.-) ; Curcumin (IT942ZTH98)
Language	English
Publishing date	2018-02-02
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms19020448
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Article ; Online: Apoptosis

Claire M. Pfeffer / Amareshwar T. K. Singh

International Journal of Molecular Sciences, Vol 19, Iss 2, p

A Target for Anticancer Therapy

2018 Volume 448

Abstract: Apoptosis, the cell’s natural mechanism for death, is a promising target for anticancer therapy. Both the intrinsic and extrinsic pathways use caspases to carry out apoptosis through the cleavage of hundreds of proteins. In cancer, the apoptotic pathway ... ...

Abstract	Apoptosis, the cell’s natural mechanism for death, is a promising target for anticancer therapy. Both the intrinsic and extrinsic pathways use caspases to carry out apoptosis through the cleavage of hundreds of proteins. In cancer, the apoptotic pathway is typically inhibited through a wide variety of means including overexpression of antiapoptotic proteins and under-expression of proapoptotic proteins. Many of these changes cause intrinsic resistance to the most common anticancer therapy, chemotherapy. Promising new anticancer therapies are plant-derived compounds that exhibit anticancer activity through activating the apoptotic pathway.
Keywords	apoptosis ; anticancer therapy ; curcumin ; apoptotic evasion ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Language	English
Publishing date	2018-02-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Genome-wide screen reveals Rab12 GTPase as a critical activator of Parkinson’s disease-linked LRRK2 kinase

Herschel S Dhekne / Francesca Tonelli / Wondwossen M Yeshaw / Claire Y Chiang / Charles Limouse / Ebsy Jaimon / Elena Purlyte / Dario R Alessi / Suzanne R Pfeffer

eLife, Vol

2023 Volume 12

Abstract: Activating mutations in the leucine-rich repeat kinase 2 (LRRK2) cause Parkinson’s disease. LRRK2 phosphorylates a subset of Rab GTPases, particularly Rab10 and Rab8A, and we showed previously that these phosphoRabs play an important role in LRRK2 ... ...

Abstract	Activating mutations in the leucine-rich repeat kinase 2 (LRRK2) cause Parkinson’s disease. LRRK2 phosphorylates a subset of Rab GTPases, particularly Rab10 and Rab8A, and we showed previously that these phosphoRabs play an important role in LRRK2 membrane recruitment and activation (Vides et al., 2022). To learn more about LRRK2 pathway regulation, we carried out an unbiased, CRISPR-based genome-wide screen to identify modifiers of cellular phosphoRab10 levels. A flow cytometry assay was developed to detect changes in phosphoRab10 levels in pools of mouse NIH-3T3 cells harboring unique CRISPR guide sequences. Multiple negative and positive regulators were identified; surprisingly, knockout of the Rab12 gene was especially effective in decreasing phosphoRab10 levels in multiple cell types and knockout mouse tissues. Rab-driven increases in phosphoRab10 were specific for Rab12, LRRK2-dependent and PPM1H phosphatase-reversible, and did not require Rab12 phosphorylation; they were seen with wild type and pathogenic G2019S and R1441C LRRK2. As expected for a protein that regulates LRRK2 activity, Rab12 also influenced primary cilia formation. AlphaFold modeling revealed a novel Rab12 binding site in the LRRK2 Armadillo domain, and we show that residues predicted to be essential for Rab12 interaction at this site influence phosphoRab10 and phosphoRab12 levels in a manner distinct from Rab29 activation of LRRK2. Our data show that Rab12 binding to a new site in the LRRK2 Armadillo domain activates LRRK2 kinase for Rab phosphorylation and could serve as a new therapeutic target for a novel class of LRRK2 inhibitors that do not target the kinase domain.
Keywords	Parkinson's disease ; Rab GTPase ; kinase ; lysosome stress ; LRRK2 ; Medicine ; R ; Science ; Q ; Biology (General) ; QH301-705.5
Subject code	571
Language	English
Publishing date	2023-10-01T00:00:00Z
Publisher	eLife Sciences Publications Ltd
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: The Evolution, Functions and Applications of the Breast Cancer Genes

Pfeffer, Claire M / Ho, Benjamin N / Singh, Amareshwar T K

Cancer genomics & proteomics

2017 Volume 14, Issue 5, Page(s) 293–298

Abstract: BRCA1 and BRCA2 are both tumor suppressors whose mutations are the cause of most hereditary breast cancers. Both genes are highly involved in ensuring genome stability. BRCA1 homologs are found in the plant and animal kingdoms while BRCA2 homologs are ... ...

Abstract	BRCA1 and BRCA2 are both tumor suppressors whose mutations are the cause of most hereditary breast cancers. Both genes are highly involved in ensuring genome stability. BRCA1 homologs are found in the plant and animal kingdoms while BRCA2 homologs are additionally found in the fungi kingdom. The initial origin of both genes remains unknown, however it is expected that the common ancestors originated around 1.6 billion years ago prior to the kingdoms diverging. There has been a great amount of divergence between homologs that is not observed in other tumor suppressors with only functionally important domains conserved. This divergence continues today with evidence of primate BRCA1/2 evolution. Cancer-associated mutations have been found to occur at conserved sites, indicating that conserved sites are important for function. In this study, we present a review on the phylogenesis of BRCA1 and BRCA2.
MeSH term(s)	BRCA1 Protein/chemistry ; BRCA1 Protein/genetics ; BRCA1 Protein/metabolism ; BRCA2 Protein/chemistry ; BRCA2 Protein/genetics ; BRCA2 Protein/metabolism ; Breast Neoplasms/genetics ; Evolution, Molecular ; Female ; Humans ; Phylogeny ; Protein Domains
Chemical Substances	BRCA1 Protein ; BRCA2 Protein
Language	English
Publishing date	2017-09-01
Publishing country	Greece
Document type	Journal Article ; Review
ZDB-ID	2144517-5
ISSN	1790-6245 ; 1109-6535
ISSN (online)	1790-6245
ISSN	1109-6535
DOI	10.21873/cgp.20040
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Zs.A 5873: Show issues

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ab Jg. 2022: Lesesaal (EG)

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Article ; Online: Update on Nanotechnology-based Drug Delivery Systems in Cancer Treatment.

Ho, Benjamin N / Pfeffer, Claire M / Singh, Amareshwar T K

Anticancer research

2017 Volume 37, Issue 11, Page(s) 5975–5981

Abstract: The emerging field of nanotechnology meets the demands for innovative approaches in the diagnosis and treatment of cancer. The nanoparticles are biocompatible and biodegradable and are made of a core, a particle that acts as a carrier, and one or more ... ...

Abstract	The emerging field of nanotechnology meets the demands for innovative approaches in the diagnosis and treatment of cancer. The nanoparticles are biocompatible and biodegradable and are made of a core, a particle that acts as a carrier, and one or more functional groups on the core which target specific sites. Nanotech in drug delivery includes nanodisks, High Density Lipoprotein nanostructures, liposomes, and gold nanoparticles. The fundamental advantages of nanoparticles are: improved delivery of water-insoluble drugs, targeted delivery, co-delivery of two or more drugs for combination therapy, and visualization of the drug delivery site by combining imaging system and a therapeutic drug. One of the potential applications of nanotechnology is in the treatment of cancer. Conventional methods for cancer treatments have included chemotherapy, surgery, or radiation. Early recognition and treatment of cancer with these approaches is still challenging. Innovative technologies are needed to overcome multidrug resistance, and increase drug localization and efficacy. Application of nanotechnology to cancer biology has brought in a new hope for developing treatment strategies on cancer. In this study, we present a review on the recent advances in nanotechnology-based approaches in cancer treatment.
MeSH term(s)	Animals ; Antineoplastic Agents/administration & dosage ; Drug Delivery Systems ; Humans ; Nanoparticles/administration & dosage ; Nanotechnology/methods ; Neoplasms/drug therapy
Chemical Substances	Antineoplastic Agents
Language	English
Publishing date	2017
Publishing country	Greece
Document type	Journal Article ; Review
ZDB-ID	604549-2
ISSN	1791-7530 ; 0250-7005
ISSN (online)	1791-7530
ISSN	0250-7005
DOI	10.21873/anticanres.12044
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Zs.A 1642: Show issues

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Article ; Online: Nonfatal Injuries Sustained in Mass Shootings in the US, 2012-2019: Injury Diagnosis Matrix, Incident Context, and Public Health Considerations.

Czaja, Matthew P / Kraus, Chadd K / Phyo, Su / Olivieri, Patrick / Mederos, Dalier R / Puente, Ivan / Mohammed, Salman / Berkeley, Ross P / Slattery, David / Gildea, Thomas H / Hardman, Claire / Palmer, Brandi / Whitmill, Melissa L / Aluyen, Una / Pinnow, Jeffery M / Young, Amanda / Eastin, Carly D / Kester, Nurani M / Works, Kaitlyn R /

Pfeffer, Andrew N / Keller, Aleksander W / Tobias, Adam / Li, Benjamin / Yorkgitis, Brian / Saadat, Soheil / Langdorf, Mark I

The western journal of emergency medicine

2023 Volume 24, Issue 3, Page(s) 552–565

Abstract: Introduction: The epidemic of gun violence in the United States (US) is exacerbated by frequent mass shootings. In 2021, there were 698 mass shootings in the US, resulting in 705 deaths and 2,830 injuries. This is a companion paper to a publication in ... ...

Abstract	Introduction: The epidemic of gun violence in the United States (US) is exacerbated by frequent mass shootings. In 2021, there were 698 mass shootings in the US, resulting in 705 deaths and 2,830 injuries. This is a companion paper to a publication in JAMA Network Open, in which the nonfatal outcomes of victims of mass shootings have been only partially described. Methods: We gathered clinical and logistic information from 31 hospitals in the US about 403 survivors of 13 mass shootings, each event involving greater than 10 injuries, from 2012-19. Local champions in emergency medicine and trauma surgery provided clinical data from electronic health records within 24 hours of a mass shooting. We organized descriptive statistics of individual-level diagnoses recorded in medical records using International Classification of Diseases codes, according to the Barell Injury Diagnosis Matrix (BIDM), a standardized tool that classifies 12 types of injuries within 36 body regions. Results: Of the 403 patients who were evaluated at a hospital, 364 sustained physical injuries-252 by gunshot wound (GSW) and 112 by non-ballistic trauma-and 39 were uninjured. Fifty patients had 75 psychiatric diagnoses. Nearly 10% of victims came to the hospital for symptoms triggered by, but not directly related to, the shooting, or for exacerbations of underlying conditions. There were 362 gunshot wounds recorded in the Barell Matrix (1.44 per patient). The Emergency Severity Index (ESI) distribution was skewed toward higher acuity than typical for an emergency department (ED), with 15.1% ESI 1 and 17.6% ESI 2 patients. Semi-automatic firearms were used in 100% of these civilian public mass shootings, with 50 total weapons for 13 shootings (Route 91 Harvest Festival, Las Vegas. 24). Assailant motivations were reported to be associated with hate crimes in 23.1%. Conclusion: Survivors of mass shootings have substantial morbidity and characteristic injury distribution, but 37% of victims had no GSW. Law enforcement, emergency medical systems, and hospital and ED disaster planners can use this information for injury mitigation and public policy planning. The BIDM is useful to organize data regarding gun violence injuries. We call for additional research funding to prevent and mitigate interpersonal firearm injuries, and for the National Violent Death Reporting System to expand tracking of injuries, their sequelae, complications, and societal costs.
MeSH term(s)	Humans ; United States/epidemiology ; Wounds, Gunshot/epidemiology ; Firearms ; Public Health ; Mental Disorders ; Mass Casualty Incidents ; Homicide
Language	English
Publishing date	2023-05-02
Publishing country	United States
Document type	Journal Article
ZDB-ID	2375700-0
ISSN	1936-9018 ; 1936-9018
ISSN (online)	1936-9018
ISSN	1936-9018
DOI	10.5811/westjem.58395
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