74 results on '"Joshua C. Snyder"'
Search Results
2. Correction: The Stem Cell-Expressed Receptor Lgr5 Possesses Canonical and Functionally Active Molecular Determinants Critical to β-arrestin-2 Recruitment.
- Author
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Joshua C. Snyder, Lauren K. Rochelle, Larry S. Barak, and Marc G. Caron
- Subjects
Medicine ,Science - Published
- 2014
- Full Text
- View/download PDF
3. Veterinary Management of Harderian Gland Tumors in Cancer Rainbow (crainbow) HER2-Positive Mice
- Author
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Angela Garner, Joshua D Ginzel, Joshua C Snyder, Jeffrey I Everitt, and Chelsea D Landon
- Subjects
Case Studies ,General Veterinary ,General Biochemistry, Genetics and Molecular Biology - Abstract
A Cancer Rainbow mouse line that expresses 3 fluorescently labeled isoforms of the tumor-driver gene HER2 (HER2BOW) was developed recently for the study of tumorigenesis in the mammary gland. The expression of 1 of the 3 HER2 isoforms in HER2BOW mice is induced through the Cre/lox system. However, in addition to developing palpable mammary tumors, HER2BOW mice developed orbital tumors, specifically of the Harderian gland. Mice were euthanized, and histopathologic examination of the Harderian gland tumors was performed. Tumors were characterized by adenomatous hyperplasia to multinodular adenomas of the Harderian gland. Fluorescent imaging of the Harderian gland tissue confirmed the expression of HER2 in the tumors. Here we discuss monitoring and palliative approaches to allow attainment of humane experimental endpoints of mammary tumor growth in this mouse line. We describe a range of interventions, including close monitoring, topical palliative care, and surgical bilateral enucleation. Based on our data and previous reports in the literature, the overexpression of HER2 in Harderian gland tissue and subsequent tumor formation likely was driven by MMTV–Cre expression in the Harderian gland.
- Published
- 2022
4. A LGR5 reporter pig model closely resembles human intestine for improved study of stem cells in disease
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Cecilia R. Schaaf, Kathryn M. Polkoff, Amber Carter, Amy S. Stewart, Breanna Sheahan, John Freund, Joshua Ginzel, Joshua C. Snyder, Jatin Roper, Jorge A. Piedrahita, and Liara M. Gonzalez
- Subjects
Genetics ,Molecular Biology ,Biochemistry ,Biotechnology - Published
- 2023
5. Supplementary Figure 7 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
mRNA FISH of cell state markers
- Published
- 2023
6. Movie 2 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
d16HER2 labelled sidebuds. d16HER2 shown in yellow. Krt14 antibody staining shown in green.
- Published
- 2023
7. Supplementary Figure 2 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
Analysis of HER2BOW tumors
- Published
- 2023
8. Movie 3 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
p95HER2 invasive carcinoma. p95HER2 shown in magenta. Krt14 antibody staining shown in green.
- Published
- 2023
9. Data from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
HER2-positive breast cancers are among the most heterogeneous breast cancer subtypes. The early amplification of HER2 and its known oncogenic isoforms provide a plausible mechanism in which distinct programs of tumor heterogeneity could be traced to the initial oncogenic event. Here a Cancer rainbow mouse simultaneously expressing fluorescently barcoded wildtype (WTHER2), exon-16 null (d16HER2), and N-terminally truncated (p95HER2) HER2 isoforms is used to trace tumorigenesis from initiation to invasion. Tumorigenesis was visualized using whole-gland fluorescent lineage tracing and single-cell molecular pathology. We demonstrate that within weeks of expression, morphologic aberrations were already present and unique to each HER2 isoform. Although WTHER2 cells were abundant throughout the mammary ducts, detectable lesions were exceptionally rare. In contrast, d16HER2 and p95HER2 induced rapid tumor development. d16HER2 incited homogenous and proliferative luminal-like lesions which infrequently progressed to invasive phenotypes whereas p95HER2 lesions were heterogenous and invasive at the smallest detectable stage. Distinct cancer trajectories were observed for d16HER2 and p95HER2 tumors as evidenced by oncogene-dependent changes in epithelial specification and the tumor microenvironment. These data provide direct experimental evidence that intratumor heterogeneity programs begin very early and well in advance of screen or clinically detectable breast cancer.Implications:Although all HER2 breast cancers are treated equally, we show a mechanism by which clinically undetected HER2 isoforms program heterogenous cancer phenotypes through biased epithelial specification and adaptations within the tumor microenvironment.
- Published
- 2023
10. Supplementary File 1 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
Supplementary File 1.
- Published
- 2023
11. Supplementary Figure 3 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
UMAP clustering of sequenced cells and cluster identification
- Published
- 2023
12. Supplementary Figure 5 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
TruRegistry of HER2BOW tissues
- Published
- 2023
13. Movie 5 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
d16HER2 mammary intraepithelial neoplasm. d16HER2 shown in magenta. Krt14 antibody staining shown in green.
- Published
- 2023
14. Movie 4 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
p95HER2 microinvasive carcinoma. p95HER2 shown in magenta. Krt14 antibody staining shown in green.
- Published
- 2023
15. Supplementary Tables 1-5 for scRNAseq marker genes from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
Supplementary Table 1: Cluster-specific markers from all cells (8,486 cells) Supplementary Table 2: Cluster-specific markers from all epithelial cells (3,843 cells) Supplementary Table 3: Cluster-specific markers from all immune cells (1,803 cells) Supplementary Table 4: Cluster-specific markers from all fibroblast cells (922 cells) Supplementary Table 5a: State-specific markers from all d16:HBOW epithelial cells (1,421 cells) Supplementary Table 5b: State-specific markers from all p95:HBOW epithelial cells (1,258 cells)
- Published
- 2023
16. Supplementary Figure 6 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
Geneset analysis of HER2BOW trajectory mapping
- Published
- 2023
17. Movie 1 from HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
- Author
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Joshua C. Snyder, H. Kim Lyerly, Alexander D. Borowsky, Robert D. Cardiff, Neil E. Hubbard, Jane Q. Chen, Marc G. Caron, Lawrence S. Barak, Erika J. Crosby, Zachary C. Hartman, Jeffrey I. Everitt, Wendy L. Roberts, Lauren K. Rochelle, Peter G. Boone, Hidetoshi Mori, Veronica Lubkov, Chaitanya R. Acharya, and Joshua D. Ginzel
- Abstract
p95HER2 labelled sidebuds. p95HER2 shown in magenta. Krt14 antibody staining shown in green.
- Published
- 2023
18. Table S1 from Vaccine-Induced Memory CD8+ T Cells Provide Clinical Benefit in HER2 Expressing Breast Cancer: A Mouse to Human Translational Study
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Zachary C. Hartman, H. Kim Lyerly, Michael A. Morse, Amy C. Hobeika, Takuya Osada, Joshua C. Snyder, Veronica Lubkov, Andre Rogatko, Sungjin Kim, Terry Hyslop, Gloria Broadwater, Holden T. Maecker, Serena Chang, Paul K. Marcom, Kimberly Blackwell, William Gwin, and Erika J. Crosby
- Abstract
CYTOF Panel
- Published
- 2023
19. Figure S3 from Vaccine-Induced Memory CD8+ T Cells Provide Clinical Benefit in HER2 Expressing Breast Cancer: A Mouse to Human Translational Study
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Zachary C. Hartman, H. Kim Lyerly, Michael A. Morse, Amy C. Hobeika, Takuya Osada, Joshua C. Snyder, Veronica Lubkov, Andre Rogatko, Sungjin Kim, Terry Hyslop, Gloria Broadwater, Holden T. Maecker, Serena Chang, Paul K. Marcom, Kimberly Blackwell, William Gwin, and Erika J. Crosby
- Abstract
SPADE tree generated by CYTOF analysis
- Published
- 2023
20. Figure S4 from Stimulation of Oncogene-Specific Tumor-Infiltrating T Cells through Combined Vaccine and αPD-1 Enable Sustained Antitumor Responses against Established HER2 Breast Cancer
- Author
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Zachary C. Hartman, Herbert K. Lyerly, Michael A. Morse, Jeremy Force, Joshua C. Snyder, Benjamin G. Vincent, Benjamin K. Ashby, Shengjie Chai, Charles M. Perou, Xiaping He, Daniel P. Hollern, Jonathan H. Shepherd, Terry Hyslop, Gloria Broadwater, Lewis A. Chodosh, William J. Muller, Kay U. Wagner, Cong-Xiao Liu, Tao Wang, Xiao-Yi Yang, Jun-Ping Wei, Gangjun Lei, Christopher A. Rabiola, Anthony-Fayez Haddad, Chaitanya R. Acharya, and Erika J. Crosby
- Abstract
Supplementary Figure 4
- Published
- 2023
21. Table S1 from Stimulation of Oncogene-Specific Tumor-Infiltrating T Cells through Combined Vaccine and αPD-1 Enable Sustained Antitumor Responses against Established HER2 Breast Cancer
- Author
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Zachary C. Hartman, Herbert K. Lyerly, Michael A. Morse, Jeremy Force, Joshua C. Snyder, Benjamin G. Vincent, Benjamin K. Ashby, Shengjie Chai, Charles M. Perou, Xiaping He, Daniel P. Hollern, Jonathan H. Shepherd, Terry Hyslop, Gloria Broadwater, Lewis A. Chodosh, William J. Muller, Kay U. Wagner, Cong-Xiao Liu, Tao Wang, Xiao-Yi Yang, Jun-Ping Wei, Gangjun Lei, Christopher A. Rabiola, Anthony-Fayez Haddad, Chaitanya R. Acharya, and Erika J. Crosby
- Abstract
Supplementary Table 1
- Published
- 2023
22. Data from Stimulation of Oncogene-Specific Tumor-Infiltrating T Cells through Combined Vaccine and αPD-1 Enable Sustained Antitumor Responses against Established HER2 Breast Cancer
- Author
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Zachary C. Hartman, Herbert K. Lyerly, Michael A. Morse, Jeremy Force, Joshua C. Snyder, Benjamin G. Vincent, Benjamin K. Ashby, Shengjie Chai, Charles M. Perou, Xiaping He, Daniel P. Hollern, Jonathan H. Shepherd, Terry Hyslop, Gloria Broadwater, Lewis A. Chodosh, William J. Muller, Kay U. Wagner, Cong-Xiao Liu, Tao Wang, Xiao-Yi Yang, Jun-Ping Wei, Gangjun Lei, Christopher A. Rabiola, Anthony-Fayez Haddad, Chaitanya R. Acharya, and Erika J. Crosby
- Abstract
Purpose:Despite promising advances in breast cancer immunotherapy, augmenting T-cell infiltration has remained a significant challenge. Although neither individual vaccines nor immune checkpoint blockade (ICB) have had broad success as monotherapies, we hypothesized that targeted vaccination against an oncogenic driver in combination with ICB could direct and enable antitumor immunity in advanced cancers.Experimental Design:Our models of HER2+ breast cancer exhibit molecular signatures that are reflective of advanced human HER2+ breast cancer, with a small numbers of neoepitopes and elevated immunosuppressive markers. Using these, we vaccinated against the oncogenic HER2Δ16 isoform, a nondriver tumor-associated gene (GFP), and specific neoepitopes. We further tested the effect of vaccination or anti–PD-1, alone and in combination.Results:We found that only vaccination targeting HER2Δ16, a driver of oncogenicity and HER2-therapeutic resistance, could elicit significant antitumor responses, while vaccines targeting a nondriver tumor-specific antigen or tumor neoepitopes did not. Vaccine-induced HER2-specific CD8+ T cells were essential for responses, which were more effective early in tumor development. Long-term tumor control of advanced cancers occurred only when HER2Δ16 vaccination was combined with αPD-1. Single-cell RNA sequencing of tumor-infiltrating T cells revealed that while vaccination expanded CD8 T cells, only the combination of vaccine with αPD-1 induced functional gene expression signatures in those CD8 T cells. Furthermore, we show that expanded clones are HER2-reactive, conclusively demonstrating the efficacy of this vaccination strategy in targeting HER2.Conclusions:Combining oncogenic driver targeted vaccines with selective ICB offers a rational paradigm for precision immunotherapy, which we are clinically evaluating in a phase II trial (NCT03632941).
- Published
- 2023
23. Supplementary methods from Stimulation of Oncogene-Specific Tumor-Infiltrating T Cells through Combined Vaccine and αPD-1 Enable Sustained Antitumor Responses against Established HER2 Breast Cancer
- Author
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Zachary C. Hartman, Herbert K. Lyerly, Michael A. Morse, Jeremy Force, Joshua C. Snyder, Benjamin G. Vincent, Benjamin K. Ashby, Shengjie Chai, Charles M. Perou, Xiaping He, Daniel P. Hollern, Jonathan H. Shepherd, Terry Hyslop, Gloria Broadwater, Lewis A. Chodosh, William J. Muller, Kay U. Wagner, Cong-Xiao Liu, Tao Wang, Xiao-Yi Yang, Jun-Ping Wei, Gangjun Lei, Christopher A. Rabiola, Anthony-Fayez Haddad, Chaitanya R. Acharya, and Erika J. Crosby
- Abstract
Supplementary methods
- Published
- 2023
24. Data from Vaccine-Induced Memory CD8+ T Cells Provide Clinical Benefit in HER2 Expressing Breast Cancer: A Mouse to Human Translational Study
- Author
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Zachary C. Hartman, H. Kim Lyerly, Michael A. Morse, Amy C. Hobeika, Takuya Osada, Joshua C. Snyder, Veronica Lubkov, Andre Rogatko, Sungjin Kim, Terry Hyslop, Gloria Broadwater, Holden T. Maecker, Serena Chang, Paul K. Marcom, Kimberly Blackwell, William Gwin, and Erika J. Crosby
- Abstract
Purpose:Immune-based therapy for metastatic breast cancer has had limited success, particularly in molecular subtypes with low somatic mutations rates. Strategies to augment T-cell infiltration of tumors include vaccines targeting established oncogenic drivers such as the genomic amplification of HER2. We constructed a vaccine based on a novel alphaviral vector encoding a portion of HER2 (VRP-HER2).Patients and Methods:In preclinical studies, mice were immunized with VRP-HER2 before or after implantation of hHER2+ tumor cells and HER2-specific immune responses and antitumor function were evaluated. We tested VRP-HER2 in a phase I clinical trial where subjects with advanced HER2-overexpressing malignancies in cohort 1 received VRP-HER2 every 2 weeks for a total of 3 doses. In cohort 2, subjects received the same schedule concurrently with a HER2-targeted therapy.Results:Vaccination in preclinical models with VRP-HER2 induced HER2-specific T cells and antibodies while inhibiting tumor growth. VRP-HER2 was well tolerated in patients and vaccination induced HER2-specific T cells and antibodies. Although a phase I study, there was 1 partial response and 2 patients with continued stable disease. Median OS was 50.2 months in cohort 1 (n = 4) and 32.7 months in cohort 2 (n = 18). Perforin expression by memory CD8 T cells post-vaccination significantly correlated with improved PFS.Conclusions:VRP-HER2 increased HER2-specific memory CD8 T cells and had antitumor effects in preclinical and clinical studies. The expansion of HER2-specific memory CD8 T cells in vaccinated patients was significantly correlated with increased PFS. Subsequent studies will seek to enhance T-cell activity by combining with anti-PD-1.
- Published
- 2023
25. HSP90-Specific nIR Probe Identifies Aggressive Prostate Cancers: Translation from Preclinical Models to a Human Phase I Study
- Author
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Chaitanya R. Acharya, Xiao Yi Yang, H. Kim Lyerly, Joshua C. Snyder, Erika J. Crosby, Michael A. Morse, Rendon C. Nelson, Thomas J. Polascik, Ivan Spasojevic, Takuya Osada, Joshua D. Ginzel, Kensuke Kaneko, Timothy A.J. Haystead, Andre Rogatko, Jiaoti Huang, Amy Hobeika, Zachary C. Hartman, Philip F. Hughes, and Leonard M. Neckers
- Subjects
Male ,Cancer Research ,medicine.medical_treatment ,Mice, SCID ,Article ,Mice ,Prostate cancer ,Prostate ,Cell Line, Tumor ,Heat shock protein ,medicine ,Animals ,Humans ,HSP90 Heat-Shock Proteins ,biology ,Prostatectomy ,business.industry ,Prostatic Neoplasms ,medicine.disease ,Hsp90 ,Epithelium ,Androgen receptor ,medicine.anatomical_structure ,Oncology ,biology.protein ,Cancer research ,business ,Erg - Abstract
A noninvasive test to discriminate indolent prostate cancers from lethal ones would focus treatment where necessary while reducing overtreatment. We exploited the known activity of heat shock protein 90 (Hsp90) as a chaperone critical for the function of numerous oncogenic drivers, including the androgen receptor and its variants, to detect aggressive prostate cancer. We linked a near-infrared fluorescing molecule to an HSP90 binding drug and demonstrated that this probe (designated HS196) was highly sensitive and specific for detecting implanted prostate cancer cell lines with greater uptake by more aggressive subtypes. In a phase I human study, systemically administered HS196 could be detected in malignant nodules within prostatectomy specimens. Single-cell RNA sequencing identified uptake of HS196 by malignant prostate epithelium from the peripheral zone (AMACR+ERG+EPCAM+ cells), including SYP+ neuroendocrine cells that are associated with therapeutic resistance and metastatic progression. A theranostic version of this molecule is under clinical testing.
- Published
- 2021
26. Integrating Clinical Trial Imaging Data Resources Using Service-Oriented Architecture and Grid Computing.
- Author
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Stefan Baumann El-Ghatta, Thierry Cladé, and Joshua C. Snyder
- Published
- 2010
- Full Text
- View/download PDF
27. Measuring Nonapoptotic Caspase Activity with a Transgenic Reporter in Mice
- Author
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P. J. Nicholls, Thomas F. Pack, Nikhil M. Urs, Sunil Kumar, Yang Zhou, Gabriel Ichim, Joshua D. Ginzel, Gabor Turu, Evan Calabrese, Wendy L. Roberts, Ping Fan, Valeriy G. Ostapchenko, Monica S. Guzman Lenis, Flavio Beraldo, Jiri Hatina, Vania F. Prado, Marco A. M. Prado, Ivan Spasojevic, Joshua C. Snyder, Kafui Dzirasa, G. Allan Johnson, Marc G. Caron, Manship, Brigitte, Duke University Medical Center, Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), University of Western Ontario (UWO), Charles University [Prague] (CU), and Duke University [Durham]
- Subjects
Genetically modified mouse ,sex differences ,Programmed cell death ,Transgene ,Cell ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Biology ,03 medical and health sciences ,stress ,0302 clinical medicine ,[SDV.CAN] Life Sciences [q-bio]/Cancer ,In vivo ,Neuroplasticity ,medicine ,mapping ,030304 developmental biology ,0303 health sciences ,nonapoptotic ,General Neuroscience ,amygdala ,in vivo reporter ,General Medicine ,Cell biology ,medicine.anatomical_structure ,caspases ,Apoptosis ,Caspase 10 ,030217 neurology & neurosurgery - Abstract
The protease caspase-3 is a key mediator of apoptotic programmed cell death. But weak or transient caspase activity can contribute to neuronal differentiation, axonal pathfinding, and synaptic long-term depression. Despite the importance of sublethal, or nonapoptotic, caspase activity in neurodevelopment and neural plasticity, there has been no simple method for mapping and quantifying nonapoptotic caspase activity in rodent brains. We therefore generated a transgenic mouse expressing a highly sensitive and specific fluorescent reporter of caspase activity, with peak signal localized to the nucleus. As a proof of concept, we first obtained evidence that nonapoptotic caspase activity influences neurophysiology in an amygdalar circuit. Then focusing on the amygdala, we were able to quantify a sex-specific persistent elevation in caspase activity in females after restraint stress. This simple in vivo caspase activity reporter will facilitate systems-level studies of apoptotic and nonapoptotic phenomena in behavioral and pathological models.Significance StatementCaspase-3 is an enzyme that can cause cell death when highly active but can also perform important cellular functions, such as maturation and structural changes, when only weakly or transiently active. Despite the importance of this nonlethal type of caspase activity, there is no straightforward method to measure it in live rodents. We therefore developed mice that have a fluorescent reporter that is sensitive enough to detect nonlethal caspase activity. Surprisingly, we found that weak caspase activity influences the synchrony of neuronal firing across different brain regions. We also observed increased caspase activity in female mice after severe stress. This simple caspase activity reporter can subserve multiple applications in behavior and pathology research.
- Published
- 2022
28. Vaccine-Induced Memory CD8+ T Cells Provide Clinical Benefit in HER2 Expressing Breast Cancer: A Mouse to Human Translational Study
- Author
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Zachary C. Hartman, Veronica Lubkov, Erika J. Crosby, Joshua C. Snyder, Sungjin Kim, Kimberly L. Blackwell, Takuya Osada, William R. Gwin, Amy Hobeika, Gloria Broadwater, Terry Hyslop, H. Kim Lyerly, Michael A. Morse, Serena Chang, Paul K. Marcom, Andre Rogatko, and Holden T. Maecker
- Subjects
0301 basic medicine ,Oncology ,Cancer Research ,medicine.medical_specialty ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Breast cancer ,Immunity ,Internal medicine ,medicine ,Cytotoxic T cell ,skin and connective tissue diseases ,neoplasms ,biology ,business.industry ,medicine.disease ,Metastatic breast cancer ,Vaccination ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cohort ,biology.protein ,Antibody ,business - Abstract
Purpose: Immune-based therapy for metastatic breast cancer has had limited success, particularly in molecular subtypes with low somatic mutations rates. Strategies to augment T-cell infiltration of tumors include vaccines targeting established oncogenic drivers such as the genomic amplification of HER2. We constructed a vaccine based on a novel alphaviral vector encoding a portion of HER2 (VRP-HER2). Patients and Methods: In preclinical studies, mice were immunized with VRP-HER2 before or after implantation of hHER2+ tumor cells and HER2-specific immune responses and antitumor function were evaluated. We tested VRP-HER2 in a phase I clinical trial where subjects with advanced HER2-overexpressing malignancies in cohort 1 received VRP-HER2 every 2 weeks for a total of 3 doses. In cohort 2, subjects received the same schedule concurrently with a HER2-targeted therapy. Results: Vaccination in preclinical models with VRP-HER2 induced HER2-specific T cells and antibodies while inhibiting tumor growth. VRP-HER2 was well tolerated in patients and vaccination induced HER2-specific T cells and antibodies. Although a phase I study, there was 1 partial response and 2 patients with continued stable disease. Median OS was 50.2 months in cohort 1 (n = 4) and 32.7 months in cohort 2 (n = 18). Perforin expression by memory CD8 T cells post-vaccination significantly correlated with improved PFS. Conclusions: VRP-HER2 increased HER2-specific memory CD8 T cells and had antitumor effects in preclinical and clinical studies. The expansion of HER2-specific memory CD8 T cells in vaccinated patients was significantly correlated with increased PFS. Subsequent studies will seek to enhance T-cell activity by combining with anti-PD-1.
- Published
- 2019
29. Slow-release delivery enhances the pharmacological properties of oral 5-hydroxytryptophan: mouse proof-of-concept
- Author
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Joshua C. Snyder, Wendy Roberts, Zixuan Yi, Nathan Modesto, Manuel Cajina, Rebecca Vernon, Peter J. Nicholls, Elizabeth L Royer, Nikhil M. Urs, Benjamin D. Sachs, Adrianna Oh, Jacob P. R. Jacobsen, Marc G. Caron, Rachel E. Bangle, and Sonora A Windermere
- Subjects
Male ,Drug ,media_common.quotation_subject ,Administration, Oral ,Mice, Transgenic ,Context (language use) ,Pharmacology ,Proof of Concept Study ,Article ,5-Hydroxytryptophan ,03 medical and health sciences ,chemistry.chemical_compound ,Therapeutic approach ,0302 clinical medicine ,Pharmacokinetics ,Fluoxetine ,medicine ,Animals ,Adverse effect ,media_common ,Brain Chemistry ,Behavior, Animal ,biology ,business.industry ,030227 psychiatry ,Psychiatry and Mental health ,chemistry ,biology.protein ,Female ,business ,Selective Serotonin Reuptake Inhibitors ,030217 neurology & neurosurgery ,medicine.drug ,Neurotrophin - Abstract
5-hydroxytryptophan (5-HTP) has shown therapeutic promise in a range of human CNS disorders. But native 5-HTP immediate release (IR) is poorly druggable, as rapid absorption causes rapid onset of adverse events, and rapid elimination causes fluctuating exposure. Recently, we reported that 5-HTP delivered as slow-release (SR) in mice augmented the brain pro-serotonergic effect of selective serotonin reuptake inhibitors (SSRIs), without the usual adverse events associated with 5-HTP IR. However, our previous study entailed translational limitations, in terms of route, dose, and duration. Here we modeled oral 5-HTP SR in mice by administering 5-HTP via the food. We modeled oral SSRI treatment via fluoxetine in the water, in a regimen recapitulating clinical pharmacokinetics and pharmacodynamics. 5-HTP SR produced plasma 5-HTP levels well within the range enhancing brain 5-HT function in humans. 5-HTP SR robustly increased brain 5-HT synthesis and levels. When administered with an SSRI, 5-HTP SR enhanced 5-HT-sensitive behaviors and neurotrophic mRNA expression. 5-HTP SR's pro-serotonergic effects were stronger in mice with endogenous brain 5-HT deficiency. In a comprehensive screen, 5-HTP SR was devoid of overt toxicological effects. The present preclinical data, appreciated in the context of published 5-HTP clinical data, suggest that 5-HTP SR could represent a new therapeutic approach to the plethora of CNS disorders potentially treatable with a pro-serotonergic drug. 5-HTP SR might in particular be therapeutically relevant when brain 5-HT deficiency is pathogenic and as an adjunctive augmentation therapy to SSRI therapy.
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- 2019
30. Abstract P2-09-16: CD8 T cells induced by novel alphaviral vector predict improved progression free survival in advanced HER2+ breast cancer patients
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Veronica Lubkov, Gloria Broadwater, Michael A. Morse, Herbert Kim Lyerly, Serena Chang, Joshua C. Snyder, Zachary C. Hartman, Terry Hyslop, Holden T. Maecker, William R. Gwin, Takuya Osada, Amy Hobeika, and Erika J. Crosby
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Oncology ,Cancer Research ,medicine.medical_specialty ,business.industry ,T cell ,Cancer ,medicine.disease ,Acquired immune system ,Metastatic breast cancer ,medicine.anatomical_structure ,Immune system ,Breast cancer ,Internal medicine ,medicine ,Cytotoxic T cell ,Progression-free survival ,skin and connective tissue diseases ,business - Abstract
Background: Immune-based therapy for metastatic breast cancer has had limited success. Strategies to augment adaptive immunity include vaccines targeting genomic amplifications like Human Epidermal Growth Factor Type 2 (HER2), an established driver of malignancy. Using a novel alphaviral vector, we constructed a vaccine encoding a portion of HER2 (VRP-HER2). Methods: In preclinical studies, mice were immunized before or after implantation of hHER2+ tumor cells and HER2-specific immune responses and anti-tumor function were assessed. We then translated this vaccine into a phase I clinical trial in which subjects with advanced HER2-overexpressing breast cancers received VRP-HER2 every 2 weeks for a total of three doses (cohort 1). In cohort 2, subjects received the same dose of VRP-HER2 along with a standard HER2 targeted therapy. Results: VRP-HER2 induced HER2-specific T cell and antibody responses while controlling tumor growth in murine models. Vaccination with VRP-HER2 was well tolerated in both patient cohorts. PFS was modest, while median OS was 50.2 months in cohort 1 and 32.7 months in cohort 2. In cohort 2, there is one partial response and two patients with continued stable disease. Vaccine induced anti-HER2 antibodies and T cells were identified. Increased perforin expression by memory CD8 T cells post vaccination significantly correlated with improved PFS. Conclusions: VRP-HER2 led to an increase in perforin expressing HER2-specific memory CD8 T cells in preclinical and clinical studies, and had profound antitumor effects in murine models. The generation of HER2-specific memory CD8 T cells was significantly correlated with increased PFS in patients. Subsequent studies will seek to enhance T cell activity by combination with anti-PD-1/PD-L1 antibodies. Citation Format: Crosby EJ, Gwin WR, Chang S, Maecker HT, Lubkov V, Snyder JC, Broadwater G, Hyslop T, Osada T, Hobeika AC, Hartman ZC, Morse MA, Lyerly HK. CD8 T cells induced by novel alphaviral vector predict improved progression free survival in advanced HER2+ breast cancer patients [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-09-16.
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- 2019
31. HER2 Isoforms Uniquely Program Intratumor Heterogeneity and Predetermine Breast Cancer Trajectories During the Occult Tumorigenic Phase
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Joshua D. Ginzel, Joshua C. Snyder, Marc G. Caron, Peter G. Boone, Veronica Lubkov, H. Kim Lyerly, Alexander D. Borowsky, Hidetoshi Mori, Erika J. Crosby, Lauren K. Rochelle, Zachary C. Hartman, Larry S. Barak, Neil E. Hubbard, Chaitanya R. Acharya, Wendy Roberts, Jeffrey I. Everitt, Robert D. Cardiff, and Jane Q. Chen
- Subjects
Gene isoform ,Cancer Research ,Carcinogenesis ,Receptor, ErbB-2 ,Knockout ,Oncology and Carcinogenesis ,Breast Neoplasms ,Biology ,medicine.disease_cause ,Article ,Mice ,Breast cancer ,ErbB-2 ,Breast Cancer ,medicine ,Tumor Microenvironment ,2.1 Biological and endogenous factors ,Animals ,Protein Isoforms ,Oncology & Carcinogenesis ,Aetiology ,skin and connective tissue diseases ,Molecular Biology ,neoplasms ,Cancer ,Mice, Knockout ,Tumor microenvironment ,Neoplastic ,Molecular pathology ,Wild type ,medicine.disease ,Phenotype ,Gene Expression Regulation, Neoplastic ,Good Health and Well Being ,Oncology ,Gene Expression Regulation ,Cancer research ,Female ,Receptor ,Developmental Biology - Abstract
HER2-positive breast cancers are among the most heterogeneous breast cancer subtypes. The early amplification of HER2 and its known oncogenic isoforms provide a plausible mechanism in which distinct programs of tumor heterogeneity could be traced to the initial oncogenic event. Here a Cancer rainbow mouse simultaneously expressing fluorescently barcoded wildtype (WTHER2), exon-16 null (d16HER2), and N-terminally truncated (p95HER2) HER2 isoforms is used to trace tumorigenesis from initiation to invasion. Tumorigenesis was visualized using whole-gland fluorescent lineage tracing and single-cell molecular pathology. We demonstrate that within weeks of expression, morphologic aberrations were already present and unique to each HER2 isoform. Although WTHER2 cells were abundant throughout the mammary ducts, detectable lesions were exceptionally rare. In contrast, d16HER2 and p95HER2 induced rapid tumor development. d16HER2 incited homogenous and proliferative luminal-like lesions which infrequently progressed to invasive phenotypes whereas p95HER2 lesions were heterogenous and invasive at the smallest detectable stage. Distinct cancer trajectories were observed for d16HER2 and p95HER2 tumors as evidenced by oncogene-dependent changes in epithelial specification and the tumor microenvironment. These data provide direct experimental evidence that intratumor heterogeneity programs begin very early and well in advance of screen or clinically detectable breast cancer. Implications: Although all HER2 breast cancers are treated equally, we show a mechanism by which clinically undetected HER2 isoforms program heterogenous cancer phenotypes through biased epithelial specification and adaptations within the tumor microenvironment.
- Published
- 2021
32. Noncanonical scaffolding of G αi and β-arrestin by G protein–coupled receptors
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Claudia Lee, Jeffrey S. Smith, Dean P. Staus, Sudarshan Rajagopal, Thomas F. Pack, Issac Choi, Xinyu Xiong, Zhiyuan Ma, Dylan Eiger, Ian M. Levitan, Gayathri Viswanathan, Alem W. Kahsai, Kevin Zheng, Joshua C. Snyder, Marc G. Caron, Anmol Warman, Asuka Inoue, and Lauren K. Rochelle
- Subjects
Multidisciplinary ,Chemistry ,G protein ,GTP-Binding Protein alpha Subunits ,Heterotrimeric G protein ,Gi alpha subunit ,Arrestin ,Signal transducing adaptor protein ,Signal transduction ,G protein-coupled receptor ,Cell biology - Abstract
Another way for GPCRs to signal G protein–coupled receptors (GPCRs) normally transmit signals by coupling to heterotrimeric guanine nucleotide–binding proteins (G proteins) or by binding β-arrestin proteins. Smith et al. provide evidence for another mechanism, an approximate combination of the two. They monitored the interaction of vasopressin type 2 receptors (V2Rs) and G α proteins in cultured cells using bioluminescent resonance energy transfer. Even though V2Rs do not signal canonically through G α i proteins, they promoted the formation of complexes containing β-arrestin and G α i , and this led to downstream signaling to extracellular signal-regulated kinase protein kinases. Science , this issue p. eaay1833
- Published
- 2021
33. Stimulation of oncogene-specific tumor infiltrating T-cells through combined vaccine and αPD1 enable sustained anti-tumor responses against established HER2 Breast Cancer
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Herbert Kim Lyerly, Daniel P. Hollern, Kay U. Wagner, Anthony-Fayez Haddad, Erika J. Crosby, William J. Muller, Chaitanya R. Acharya, Gloria Broadwater, Michael A. Morse, Xiaping He, Junping Wei, Shengjie Chai, Charles M. Perou, Lewis A. Chodosh, Tao Wang, Terry Hyslop, Joshua C. Snyder, Gangjun Lei, Benjamin G. Vincent, Jonathan Shepherd, Christopher A. Rabiola, Cong-Xiao Liu, Jeremy Force, Benjamin K. Ashby, Zachary C. Hartman, and Xiao Yi Yang
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0301 basic medicine ,Cancer Research ,Receptor, ErbB-2 ,medicine.medical_treatment ,Programmed Cell Death 1 Receptor ,Breast Neoplasms ,Mice, Transgenic ,CD8-Positive T-Lymphocytes ,Cancer Vaccines ,Article ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Breast cancer ,Lymphocytes, Tumor-Infiltrating ,Antigen ,Antigens, Neoplasm ,Cell Line, Tumor ,medicine ,Cytotoxic T cell ,Animals ,Humans ,Vaccines, Combined ,Immune Checkpoint Inhibitors ,Oncogene ,business.industry ,Mammary Neoplasms, Experimental ,Immunotherapy ,medicine.disease ,Immune checkpoint ,3. Good health ,Vaccination ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Female ,business ,CD8 - Abstract
Purpose: Despite promising advances in breast cancer immunotherapy, augmenting T-cell infiltration has remained a significant challenge. Although neither individual vaccines nor immune checkpoint blockade (ICB) have had broad success as monotherapies, we hypothesized that targeted vaccination against an oncogenic driver in combination with ICB could direct and enable antitumor immunity in advanced cancers. Experimental Design: Our models of HER2+ breast cancer exhibit molecular signatures that are reflective of advanced human HER2+ breast cancer, with a small numbers of neoepitopes and elevated immunosuppressive markers. Using these, we vaccinated against the oncogenic HER2Δ16 isoform, a nondriver tumor-associated gene (GFP), and specific neoepitopes. We further tested the effect of vaccination or anti–PD-1, alone and in combination. Results: We found that only vaccination targeting HER2Δ16, a driver of oncogenicity and HER2-therapeutic resistance, could elicit significant antitumor responses, while vaccines targeting a nondriver tumor-specific antigen or tumor neoepitopes did not. Vaccine-induced HER2-specific CD8+ T cells were essential for responses, which were more effective early in tumor development. Long-term tumor control of advanced cancers occurred only when HER2Δ16 vaccination was combined with αPD-1. Single-cell RNA sequencing of tumor-infiltrating T cells revealed that while vaccination expanded CD8 T cells, only the combination of vaccine with αPD-1 induced functional gene expression signatures in those CD8 T cells. Furthermore, we show that expanded clones are HER2-reactive, conclusively demonstrating the efficacy of this vaccination strategy in targeting HER2. Conclusions: Combining oncogenic driver targeted vaccines with selective ICB offers a rational paradigm for precision immunotherapy, which we are clinically evaluating in a phase II trial (NCT03632941).
- Published
- 2020
34. Heat shock protein 90-targeted photodynamic therapy enables treatment of subcutaneous and visceral tumors
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H. Kim Lyerly, Timothy Aj. Haystead, Joshua D. Ginzel, Joshua C. Snyder, Kensuke Kaneko, Xiao Yi Yang, Michael A. Morse, Takuya Osada, Cong-Xiao Liu, Philip F. Hughes, William R. Gwin, Marcio A. Diniz, and Khaldon Bodoor
- Subjects
medicine.medical_treatment ,Medicine (miscellaneous) ,Antineoplastic Agents ,Photodynamic therapy ,Inflammatory breast cancer ,Article ,General Biochemistry, Genetics and Molecular Biology ,Hsp90 inhibitor ,03 medical and health sciences ,Breast cancer ,Targeted therapies ,0302 clinical medicine ,Cell Line, Tumor ,polycyclic compounds ,medicine ,Humans ,Photosensitizer ,HSP90 Heat-Shock Proteins ,lcsh:QH301-705.5 ,030304 developmental biology ,0303 health sciences ,Photosensitizing Agents ,business.industry ,Verteporfin ,Cancer ,medicine.disease ,lcsh:Biology (General) ,Photochemotherapy ,030220 oncology & carcinogenesis ,Cancer cell ,MCF-7 Cells ,Cancer research ,General Agricultural and Biological Sciences ,business ,medicine.drug - Abstract
Photodynamic therapy (PDT) ablates malignancies by applying focused near-infrared (nIR) light onto a lesion of interest after systemic administration of a photosensitizer (PS); however, the accumulation of existing PS is not tumor-exclusive. We developed a tumor-localizing strategy for PDT, exploiting the high expression of heat shock protein 90 (Hsp90) in cancer cells to retain high concentrations of PS by tethering a small molecule Hsp90 inhibitor to a PS (verteporfin, VP) to create an Hsp90-targeted PS (HS201). HS201 accumulates to a greater extent than VP in breast cancer cells both in vitro and in vivo, resulting in increased treatment efficacy of HS201-PDT in various human breast cancer xenografts regardless of molecular and clinical subtypes. The therapeutic index achieved with Hsp90-targeted PDT would permit treatment not only of localized tumors, but also more diffusely infiltrating processes such as inflammatory breast cancer., Heat shock protein 90 (Hsp90) is highly expressed in cancer cells. Kaneko et al show that targeting Hsp90 on the cancer cell surface with a photosensitizer tethered to an Hsp90 small molecule inhibitor improves the efficacy of photodynamic therapy in various human breast cancer xenografts, suggesting a therapeutic avenue for human disease.
- Published
- 2020
35. A cancer rainbow mouse for visualizing the functional genomics of oncogenic clonal expansion
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H. Kim Lyerly, Marc G. Caron, Veronica Lubkov, Alexander D. Borowsky, Mei Lang Flowers, Barry R. Stripp, Wendy Roberts, Cheryl B. Bock, Peter J. Nicholls, Lauren K. Rochelle, Pankaj K. Agarwal, Larry S. Barak, Robert D. Cardiff, Peter G. Boone, Joshua C. Snyder, Joshua D. Ginzel, and Richard J. von Furstenberg
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0301 basic medicine ,Somatic cell ,Carcinogenesis ,General Physics and Astronomy ,medicine.disease_cause ,Mice ,0302 clinical medicine ,2.1 Biological and endogenous factors ,Aetiology ,lcsh:Science ,Cancer genetics ,Cancer ,Mutation ,Multidisciplinary ,Functional genomics ,3. Good health ,Mechanisms of disease ,030220 oncology & carcinogenesis ,Colonic Neoplasms ,Neoplastic Stem Cells ,Stem Cell Research - Nonembryonic - Non-Human ,Stem cell ,Science ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,medicine ,Genetics ,Animals ,Humans ,Cell Proliferation ,RSPO3 ,Animal ,General Chemistry ,Oncogenes ,medicine.disease ,Stem Cell Research ,Disease Models, Animal ,030104 developmental biology ,Disease Models ,Cancer research ,Field cancerization ,lcsh:Q ,Thrombospondins ,Digestive Diseases - Abstract
Field cancerization is a premalignant process marked by clones of oncogenic mutations spreading through the epithelium. The timescales of intestinal field cancerization can be variable and the mechanisms driving the rapid spread of oncogenic clones are unknown. Here we use a Cancer rainbow (Crainbow) modelling system for fluorescently barcoding somatic mutations and directly visualizing the clonal expansion and spread of oncogenes. Crainbow shows that mutations of ß-catenin (Ctnnb1) within the intestinal stem cell results in widespread expansion of oncogenes during perinatal development but not in adults. In contrast, mutations that extrinsically disrupt the stem cell microenvironment can spread in adult intestine without delay. We observe the rapid spread of premalignant clones in Crainbow mice expressing oncogenic Rspondin-3 (RSPO3), which occurs by increasing crypt fission and inhibiting crypt fixation. Crainbow modelling provides insight into how somatic mutations rapidly spread and a plausible mechanism for predetermining the intratumor heterogeneity found in colon cancers., Pre-malignant cells harbouring oncogenic mutations can populate and spread throughout a tissue. Here, using a rainbow mouse system, the authors explore how clonal expansion in the mouse intestine might explain high levels of intra-tumoural heterogeneity observed in the disease.
- Published
- 2019
36. hCALCRL mutation causes autosomal recessive nonimmune hydrops fetalis with lymphatic dysplasia
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Fuad Al Mutairi, Daniel O. Kechele, John Simms, Natalie R Nielsen, Reema B. Davis, Joshua C. Snyder, Harvey J. Kliman, Marc G. Caron, Duncan I. Mackie, David R. Poyner, Jonathan S. Berg, and Kathleen M. Caron
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0301 basic medicine ,Candidate gene ,Receptor activity-modifying protein ,Immunology ,Heterozygote advantage ,CALCRL ,Biology ,medicine.disease ,3. Good health ,03 medical and health sciences ,030104 developmental biology ,Lymphatic system ,RAMP2 ,Hydrops fetalis ,medicine ,Cancer research ,Immunology and Allergy ,Signal transduction - Abstract
We report the first case of nonimmune hydrops fetalis (NIHF) associated with a recessive, in-frame deletion of V205 in the G protein–coupled receptor, Calcitonin Receptor-Like Receptor (hCALCRL). Homozygosity results in fetal demise from hydrops fetalis, while heterozygosity in females is associated with spontaneous miscarriage and subfertility. Using molecular dynamic modeling and in vitro biochemical assays, we show that the hCLR(V205del) mutant results in misfolding of the first extracellular loop, reducing association with its requisite receptor chaperone, receptor activity modifying protein (RAMP), translocation to the plasma membrane and signaling. Using three independent genetic mouse models we establish that the adrenomedullin–CLR–RAMP2 axis is both necessary and sufficient for driving lymphatic vascular proliferation. Genetic ablation of either lymphatic endothelial Calcrl or nonendothelial Ramp2 leads to severe NIHF with embryonic demise and placental pathologies, similar to that observed in humans. Our results highlight a novel candidate gene for human congenital NIHF and provide structure–function insights of this signaling axis for human physiology.
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- 2018
37. Isoforms of GPCR proteins combine for diverse signalling
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Sudarshan Rajagopal and Joshua C. Snyder
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0301 basic medicine ,Gene isoform ,Multidisciplinary ,Biology ,environment and public health ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Signalling ,Drug development ,biological phenomena, cell phenomena, and immunity ,Differential expression ,Receptor ,hormones, hormone substitutes, and hormone antagonists ,030217 neurology & neurosurgery ,G protein-coupled receptor - Abstract
Many receptor proteins of the GPCR family exist in multiple isoforms. A comprehensive analysis of different combinations of GPCR isoforms that produce diverse signalling patterns in cells has implications for drug development. Differential expression of GPCR isoform combinations alters signalling.
- Published
- 2020
38. Inhibiting clathrin-mediated endocytosis of the leucine-rich G protein-coupled receptor-5 diminishes cell fitness
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Alan S. Waggoner, Lauren K. Rochelle, Larry S. Barak, Veronica Lubkov, Marc G. Caron, Caroline A. Ray, Cheryl B. Bock, Thomas F. Pack, Joshua C. Snyder, and H. Kim Lyerly
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0301 basic medicine ,media_common.quotation_subject ,education ,Biology ,Cell fate determination ,Endocytosis ,Biochemistry ,Clathrin ,Epithelium ,Lignans ,Receptors, G-Protein-Coupled ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Leucine ,Cell Line, Tumor ,Animals ,Homeostasis ,Humans ,Protein Isoforms ,Cell Lineage ,Internalization ,Wnt Signaling Pathway ,Molecular Biology ,Cell Proliferation ,media_common ,G protein-coupled receptor ,Adenosine Triphosphatases ,Stochastic Processes ,Cell growth ,Stem Cells ,Wnt signaling pathway ,Dioxolanes ,Cell Biology ,Receptor-mediated endocytosis ,Rats ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,biology.protein ,Female ,030217 neurology & neurosurgery - Abstract
The leucine-rich G protein-coupled receptor-5 (LGR5) is expressed in adult tissue stem cells of many epithelia, and its overexpression is negatively correlated with cancer prognosis. LGR5 potentiates WNT/β-catenin signaling through its unique constitutive internalization property that clears negative regulators of the WNT-receptor complex from the membrane. However, both the mechanism and physiological relevance of LGR5 internalization are unclear. Therefore, a natural product library was screened to discover LGR5 internalization inhibitors and gain mechanistic insight into LGR5 internalization. The plant lignan justicidin B blocked the constitutive internalization of LGR5. Justicidin B is structurally similar to more potent vacuolar-type H+-ATPase inhibitors, which all inhibited LGR5 internalization by blocking clathrin-mediated endocytosis. We then tested the physiological relevance of LGR5 internalization blockade in vivo. A LGR5-rainbow (LBOW) mouse line was engineered to express three different LGR5 isoforms along with unique fluorescent protein lineage reporters in the same mouse. In this manner, the effects of each isoform on cell fate can be simultaneously assessed through simple fluorescent imaging for each lineage reporter. LBOW mice express three different forms of LGR5, a wild-type form that constitutively internalizes and two mutant forms whose internalization properties have been compromised by genetic perturbations within the carboxyl-terminal tail. LBOW was activated in the intestinal epithelium, and a year-long lineage-tracing course revealed that genetic blockade of LGR5 internalization diminished cell fitness. Together these data provide proof-of-concept genetic evidence that blocking the clathrin-mediated endocytosis of LGR5 could be used to pharmacologically control cell behavior.
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- 2017
39. Noncanonical scaffolding of G
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Jeffrey S, Smith, Thomas F, Pack, Asuka, Inoue, Claudia, Lee, Kevin, Zheng, Issac, Choi, Dylan S, Eiger, Anmol, Warman, Xinyu, Xiong, Zhiyuan, Ma, Gayathri, Viswanathan, Ian M, Levitan, Lauren K, Rochelle, Dean P, Staus, Joshua C, Snyder, Alem W, Kahsai, Marc G, Caron, and Sudarshan, Rajagopal
- Subjects
Bioluminescence Resonance Energy Transfer Techniques ,HEK293 Cells ,genetic structures ,Cell Movement ,Humans ,sense organs ,GTP-Binding Protein alpha Subunits, Gi-Go ,Extracellular Signal-Regulated MAP Kinases ,beta-Arrestins ,Article ,Receptors, G-Protein-Coupled ,Signal Transduction - Abstract
Heterotrimeric guanine nucleotide–binding protein (G protein)–coupled receptors (GPCRs) are common drug targets and canonically couple to specific G(α) protein subtypes and β-arrestin adaptor proteins. G protein–mediated signaling and β-arrestin–mediated signaling have been considered separable. We show here that GPCRs promote a direct interaction between G(αi) protein subtype family members and β-arrestins regardless of their canonical G(αi) protein subtype coupling. G(αi):β-arrestin complexes bound extracellular signal-regulated kinase (ERK), and their disruption impaired both ERK activation and cell migration, which is consistent with β-arrestins requiring a functional interaction with G(αi) for certain signaling events. These results introduce a GPCR signaling mechanism distinct from canonical G protein activation in which GPCRs cause the formation of G(αi):β-arrestin signaling complexes.
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- 2019
40. Noncanonical scaffolding of Gαi and β-arrestin by G protein-coupled receptors
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Asuka Inoue, Ian M. Levitan, Jeffrey S. Smith, Issac Choi, Alem W. Kahsai, Joshua C. Snyder, Xinyu Xiong, Zhiyuan Ma, Claudia Lee, Sudarshan Rajagopal, Marc G. Caron, Kevin Zheng, Dean P. Staus, Thomas F. Pack, and Lauren K. Rochelle
- Subjects
0303 health sciences ,Gs alpha subunit ,biology ,Protein family ,G protein ,Kinase ,Chemistry ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,Gq alpha subunit ,Arrestin ,biology.protein ,Receptor ,030217 neurology & neurosurgery ,030304 developmental biology ,G protein-coupled receptor - Abstract
SummaryG-protein-coupled receptors (GPCRs) enable cells to sense and respond appropriately to hormonal and environmental signals, and are a target of ~30% of all FDA-approved medications. Canonically, each GPCR couples to distinct Gαproteins, such as Gαs, Gαi, Gαqor Gα12/13, as well as β-arrestins. These transducer proteins translate and integrate extracellular stimuli sensed by GPCRs into intracellular signals through what are broadly considered separable signalling pathways. However, the ability of Gαproteins to directly interact with β-arrestins to integrate signalling has not previously been appreciated. Here we show a novel interaction between Gαiprotein family members and β-arrestin. Gαi:β-arrestin complexes were formed by all GPCRs tested, regardless of their canonical G protein isoform coupling, and could bind both GPCRs as well as the extracellular signal-regulated kinase (ERK). This novel paradigm of Gαi:β-arrestin scaffolds enhances our understanding of GPCR signalling.
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- 2019
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41. Vaccine-Induced Memory CD8
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Erika J, Crosby, William, Gwin, Kimberly, Blackwell, Paul K, Marcom, Serena, Chang, Holden T, Maecker, Gloria, Broadwater, Terry, Hyslop, Sungjin, Kim, Andre, Rogatko, Veronica, Lubkov, Joshua C, Snyder, Takuya, Osada, Amy C, Hobeika, Michael A, Morse, H Kim, Lyerly, and Zachary C, Hartman
- Subjects
Adult ,Male ,Receptor, ErbB-2 ,Genetic Vectors ,Mice, Nude ,Apoptosis ,Breast Neoplasms ,Alphavirus ,CD8-Positive T-Lymphocytes ,Cancer Vaccines ,Article ,Mice ,Tumor Cells, Cultured ,Animals ,Humans ,skin and connective tissue diseases ,neoplasms ,Aged ,Cell Proliferation ,Mice, Inbred BALB C ,Dendritic Cells ,Middle Aged ,Prognosis ,Xenograft Model Antitumor Assays ,Immunity, Humoral ,Survival Rate ,Vaccines, Subunit ,Female ,Immunologic Memory ,Follow-Up Studies - Abstract
BACKGROUND: Immune-based therapy for metastatic breast cancer has had limited success, particularly in molecular subtypes with low somatic mutations rates. Strategies to augment T cell infiltration of tumors include vaccines targeting established oncogenic drivers like the genomic amplification of HER2. We constructed a vaccine based on a novel alphaviral vector encoding a portion of HER2 (VRP-HER2). METHODS: In preclinical studies, mice were immunized with VRP-HER2 before or after implantation of hHER2+ tumor cells and HER2-specific immune responses and anti-tumor function were evaluated. We tested VRP-HER2 in a Phase I clinical trial where subjects with advanced HER2-overexpressing malignancies in cohort 1 received VRP-HER2 every 2 weeks for a total of three doses. In cohort 2, subjects received the same schedule concurrently with a HER2-targeted therapy. RESULTS: Vaccination in preclinical models with VRP-HER2 induced HER2-specific T cells and antibodies while inhibiting tumor growth. VRP-HER2 was well tolerated in patients and vaccination induced HER2-specific T cells and antibodies. Although a phase I study, there was one partial response and two patients with continued stable disease. Median OS was 50.2 months in cohort 1 (n=4) and 32.7 months in cohort 2 (n=18). Perforin expression by memory CD8 T cells post-vaccination significantly correlated with improved PFS. CONCLUSIONS: VRP-HER2 increased HER2-specific memory CD8 T cells and had anti-tumor effects in preclinical and clinical studies. The expansion of HER2-specific memory CD8 T cells in vaccinated patients was significantly correlated with increased PFS. Subsequent studies will seek to enhance T cell activity by combining with anti-PD-1.
- Published
- 2018
42. Distinct cortical and striatal actions of a β-arrestin–biased dopamine D2 receptor ligand reveal unique antipsychotic-like properties
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Tama Evron, Patricio O'Donnell, Joshua C. Snyder, Marc G. Caron, Thomas F. Pack, Steven M. Gee, Emiliana Borrelli, Ramona M. Rodriguiz, John D. McCorvy, Xiaobao Yang, Bryan L. Roth, Nikhil M. Urs, Jian Jin, and William C. Wetsel
- Subjects
Male ,0301 basic medicine ,Mice, 129 Strain ,G-Protein-Coupled Receptor Kinase 2 ,Phencyclidine ,Biology ,Ligands ,Partial agonist ,Mice ,03 medical and health sciences ,0302 clinical medicine ,GTP-Binding Proteins ,Interneurons ,Dopamine ,Dopamine receptor D2 ,medicine ,Arrestin ,Animals ,Humans ,Receptor ,G protein-coupled receptor ,Cerebral Cortex ,Mice, Knockout ,Multidisciplinary ,Behavior, Animal ,Receptors, Dopamine D2 ,beta-Arrestin 2 ,Corpus Striatum ,Mice, Inbred C57BL ,Dopamine D2 Receptor Antagonists ,HEK293 Cells ,030104 developmental biology ,medicine.anatomical_structure ,PNAS Plus ,Cerebral cortex ,Female ,Signal transduction ,Neuroscience ,030217 neurology & neurosurgery ,Antipsychotic Agents ,Signal Transduction ,medicine.drug - Abstract
The current dopamine (DA) hypothesis of schizophrenia postulates striatal hyperdopaminergia and cortical hypodopaminergia. Although partial agonists at DA D2 receptors (D2Rs), like aripiprazole, were developed to simultaneously target both phenomena, they do not effectively improve cortical dysfunction. In this study, we investigate the potential for newly developed β-arrestin2 (βarr2)-biased D2R partial agonists to simultaneously target hyper- and hypodopaminergia. Using neuron-specific βarr2-KO mice, we show that the antipsychotic-like effects of a βarr2-biased D2R ligand are driven through both striatal antagonism and cortical agonism of D2R-βarr2 signaling. Furthermore, βarr2-biased D2R agonism enhances firing of cortical fast-spiking interneurons. This enhanced cortical agonism of the biased ligand can be attributed to a lack of G-protein signaling and elevated expression of βarr2 and G protein-coupled receptor (GPCR) kinase 2 in the cortex versus the striatum. Therefore, we propose that βarr2-biased D2R ligands that exert region-selective actions could provide a path to develop more effective antipsychotic therapies.
- Published
- 2016
43. Constitutive Internalization of the Leucine-rich G Protein-coupled Receptor-5 (LGR5) to the Trans-Golgi Network
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Marc G. Caron, H. Kim Lyerly, Lauren K. Rochelle, Larry S. Barak, and Joshua C. Snyder
- Subjects
Time Factors ,Endosome ,Vesicular Transport Proteins ,Endosomes ,Biology ,Biochemistry ,Receptors, G-Protein-Coupled ,EEA1 ,symbols.namesake ,Humans ,Wnt Signaling Pathway ,Molecular Biology ,rab5 GTP-Binding Proteins ,G protein-coupled receptor ,Wnt signaling pathway ,LGR5 ,rab7 GTP-Binding Proteins ,Cell Biology ,Golgi apparatus ,Cell biology ,Transport protein ,Retromer complex ,Protein Transport ,HEK293 Cells ,rab GTP-Binding Proteins ,symbols ,trans-Golgi Network - Abstract
LGR5 is a Wnt pathway associated G protein-coupled receptor (GPCR) that serves as a molecular determinant of stem cells in numerous tissues including the intestine, stomach, hair follicle, eye, and mammary gland. Despite its importance as a marker for this critical niche, little is known about LGR5 signaling nor the biochemical mechanisms and receptor determinants that regulate LGR5 membrane expression and intracellular trafficking. Most importantly, in cells LGR5 is predominantly intracellular, yet the mechanisms underlying this behavior have not been determined. In this work we elucidate a precise trafficking program for LGR5 and identify the motif at its C terminus that is responsible for the observed constitutive internalization. We show that this process is dependent upon dynamin GTPase activity and find that wild-type full-length LGR5 rapidly internalizes into EEA1- and Rab5-positive endosomes. However, LGR5 fails to rapidly recycle to the plasmid membrane through Rab4-positive vesicles, as is common for other GPCRs. Rather, internalized LGR5 transits through Rab7- and Rab9-positive vesicles, co-localizes in vesicles with Vps26, a retromer complex component that regulates retrograde trafficking to the trans-Golgi network (TGN) and reaches a steady-state distribution in the TGN within 2 h. Using mutagenesis, particularly of putative phosphorylation sites, we show that the amino acid pair, serine 861 and 864, is the principal C-tail determinant that mediates LGR5 constitutive internalization. The constitutive internalization of LGR5 to the TGN suggests the existence of novel biochemical roles for its Wnt pathway related, but ill defined signaling program.
- Published
- 2013
44. Deletion of GSK3β in D2R-expressing neurons reveals distinct roles for β-arrestin signaling in antipsychotic and lithium action
- Author
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Sean M. Peterson, Jacob P. R. Jacobsen, Marc G. Caron, Nikhil M. Urs, and Joshua C. Snyder
- Subjects
Mice, 129 Strain ,Arrestins ,Aripiprazole ,Lithium ,Motor Activity ,Quinolones ,Pharmacology ,Piperazines ,Gene Knockout Techniques ,Glycogen Synthase Kinase 3 ,Mice ,GSK-3 ,Dopamine receptor D2 ,Haloperidol ,medicine ,Animals ,GSK3B ,Protein kinase B ,beta Catenin ,beta-Arrestins ,Mice, Knockout ,Glycogen Synthase Kinase 3 beta ,Multidisciplinary ,Behavior, Animal ,Receptors, Dopamine D2 ,Chemistry ,Beta-Arrestins ,Dopaminergic Neurons ,Biological Sciences ,Tail suspension test ,Mice, Inbred C57BL ,Signal transduction ,Antipsychotic Agents ,Signal Transduction ,medicine.drug - Abstract
Several studies in rodent models have shown that glycogen synthase kinase 3 β (GSK3β) plays an important role in the actions of antispychotics and mood stabilizers. Recently it was demonstrated that GSK3β through a β-arrestin2/protein kinase B (PKB or Akt)/protein phosphatase 2A (PP2A) signaling complex regulates dopamine (DA)- and lithium-sensitive behaviors and is required to mediate endophenotypes of mania and depression in rodents. We have previously shown that atypical antipsychotics antagonize DA D2 receptor (D2R)/β-arrestin2 interactions more efficaciously than G-protein–dependent signaling, whereas typical antipsychotics inhibit both pathways with similar efficacy. To elucidate the site of action of GSK3β in regulating DA- or lithium-sensitive behaviors, we generated conditional knockouts of GSK3β, where GSK3β was deleted in either DA D1- or D2-receptor–expressing neurons. We analyzed these mice for behaviors commonly used to test antipsychotic efficacy or behaviors that are sensitive to lithium treatment. Mice with deletion of GSK3β in D2 (D2GSK3β −/− ) but not D1 (D1GSK3β −/− ) neurons mimic antipsychotic action. However, haloperidol (HAL)-induced catalepsy was unchanged in either D2GSK3β −/− or D1GSK3β −/− mice compared with control mice. Interestingly, genetic stabilization of β-catenin, a downstream target of GSK3β, in D2 neurons did not affect any of the behaviors tested. Moreover, D2GSK3β −/− or D1GSK3β −/− mice showed similar responses to controls in the tail suspension test (TST) and dark–light emergence test, behaviors which were previously shown to be β-arrestin2- and GSK3β-dependent and sensitive to lithium treatment. Taken together these studies suggest that selective deletion of GSK3β but not stabilization of β-catenin in D2 neurons mimics antipsychotic action without affecting signaling pathways involved in catalepsy or certain mood-related behaviors.
- Published
- 2012
45. Chapter One - Ubiquitination and Deubiquitination of G Protein-Coupled Receptors
- Author
-
Sudha K. Shenoy, Joshua C. Snyder, and Pierre-Yves Jean-Charles
- Subjects
0301 basic medicine ,biology ,Context (language use) ,Ubiquitin ligase ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,Membrane protein ,Proteasome ,Ubiquitin ,biology.protein ,Signal transduction ,hormones, hormone substitutes, and hormone antagonists ,G protein-coupled receptor ,Deubiquitination - Abstract
The seven-transmembrane containing G protein-coupled receptors (GPCRs) constitute the largest family of cell-surface receptors. Transmembrane signaling by GPCRs is fundamental to many aspects of physiology including vision, olfaction, cardiovascular, and reproductive functions as well as pain, behavior and psychomotor responses. The duration and magnitude of signal transduction is tightly controlled by a series of coordinated trafficking events that regulate the cell-surface expression of GPCRs at the plasma membrane. Moreover, the intracellular trafficking profiles of GPCRs can correlate with the signaling efficacy and efficiency triggered by the extracellular stimuli that activate GPCRs. Of the various molecular mechanisms that impart selectivity, sensitivity and strength of transmembrane signaling, ubiquitination of the receptor protein plays an important role because it defines both trafficking and signaling properties of the activated GPCR. Ubiquitination of proteins was originally discovered in the context of lysosome-independent degradation of cytosolic proteins by the 26S proteasome; however a large body of work suggests that ubiquitination also orchestrates the downregulation of membrane proteins in the lysosomes. In the case of GPCRs, such ubiquitin-mediated lysosomal degradation engenders long-term desensitization of transmembrane signaling. To date about 40 GPCRs are known to be ubiquitinated. For many GPCRs, ubiquitination plays a major role in postendocytic trafficking and sorting to the lysosomes. This chapter will focus on the patterns and functional roles of GPCR ubiquitination, and will describe various molecular mechanisms involved in GPCR ubiquitination.
- Published
- 2016
46. Reparative Capacity of Airway Epithelium Impacts Deposition and Remodeling of Extracellular Matrix
- Author
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Anna C. Zemke, Joshua C. Snyder, and Barry R. Stripp
- Subjects
Male ,Pulmonary and Respiratory Medicine ,Pathology ,medicine.medical_specialty ,Mesenchyme ,Clinical Biochemistry ,Mitosis ,Bronchi ,Mice, Transgenic ,Naphthalenes ,Biology ,Antiviral Agents ,Epithelium ,Extracellular matrix ,Mice ,Fibrosis ,Gene expression ,medicine ,Animals ,Ganciclovir ,Lung ,Molecular Biology ,Oligonucleotide Array Sequence Analysis ,Basement membrane ,Tenascin C ,Epithelial Cells ,Articles ,Cell Biology ,respiratory system ,medicine.disease ,Extracellular Matrix ,Cell biology ,medicine.anatomical_structure ,biology.protein ,RNA ,Respiratory epithelium - Abstract
Defective epithelial repair in the setting of chronic lung disease has been suggested to contribute to uncontrolled extracellular matrix (ECM) deposition and development of fibrosis. We sought to directly test this hypothesis through gene expression profiling of total lung RNA isolated from mouse models of selective epithelial cell injury that are associated with either productive or abortive repair. Analysis of gene expression in repairing lungs of naphthalene-exposed mice revealed prominent clusters of up-regulated genes with putative roles in regulation of the extracellular matrix and cellular proliferation. Further analysis of tenascin C (Tnc), a representative matrix protein, in total lung RNA revealed a transient 4.5-fold increase in mRNA abundance 1 day after injury and a return to steady-state levels by Recovery Day 3. Tnc was deposited by the peribronchiolar mesenchyme immediately after injury and was remodeled to basement membrane subtending the bronchiolar epithelium during epithelial repair. Epithelial restitution was accompanied by a decrease in Tnc mRNA and protein expression to steady-state levels. In contrast, abortive repair using a transgenic model allowing ablation of all reparative cells led to a progressive increase in Tnc mRNA within lung tissue and accumulation of its gene product within the subepithelial mesenchyme of both conducting airways and alveoli. These data demonstrate that the ECM is dynamically remodeled in response to selective epithelial cell injury and that this process is activated without resolution in the setting of defective airway epithelial repair.
- Published
- 2009
47. Molecular Staging of Epithelial Maturation Using Secretory Cell–Specific Genes as Markers
- Author
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Susan D. Reynolds, Brian Brockway, Naftali Kaminski, Barry R. Stripp, Joshua C. Snyder, Jeffrey A. Drake, and Anna C. Zemke
- Subjects
Male ,Pulmonary and Respiratory Medicine ,Bronchiole ,Transgene ,Molecular Sequence Data ,Clinical Biochemistry ,Mice, Transgenic ,Respiratory Mucosa ,Naphthalenes ,Biology ,Epithelium ,Mice ,medicine ,Animals ,Claudin ,Lung ,Molecular Biology ,Gene ,Messenger RNA ,Gene Expression Profiling ,Membrane Proteins ,Cell Differentiation ,Epithelial Cells ,Articles ,Cell Biology ,respiratory system ,Microarray Analysis ,Embryonic stem cell ,Molecular biology ,Phenotype ,Cell biology ,medicine.anatomical_structure ,Claudins ,Biomarkers - Abstract
Bronchiolar Clara cells undergo phenotypic changes during development and in disease. These changes are poorly described due to a paucity of molecular markers. We used chemical and transgenic approaches to ablate Clara cells, allowing identification of their unique gene expression profile. Flavin monooxygenase 3 (Fmo3), paraoxonase 1 (Pon1), aldehyde oxidase 3 (Aox3), and claudin 10 (Cldn10) were identified as novel Clara cell markers. New and existing Clara cell marker genes were categorized into three classes based on their unique developmental expression pattern. Cldn10 was uniformly expressed in the epithelium at Embryonic Day (E)14.5 and became restricted to secretory cells at E18.5. This transition was defined by induction of CCSP. Maturation of secretory cells was associated with progressive increases in the expression of Fmo3, Pon1, Aox3, and Cyp2f2 between late embryonic and postnatal periods. Messenger RNA abundance of all categories of genes was dramatically decreased after naphthalene-induced airway injury, and displayed a sequence of temporal induction during repair that suggested sequential secretory cell maturation. We have defined a broader repertoire of Clara cell–specific genes that allows staging of epithelial maturation during development and repair.
- Published
- 2009
48. Endogenous lung stem cells and contribution to disease
- Author
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Barry R. Stripp, Joshua C. Snyder, and Roxana M. Teisanu
- Subjects
Lung ,Respiratory Tract Diseases ,Epithelial Cells ,Respiratory Mucosa ,respiratory system ,Biology ,Article ,Respiratory Tract Neoplasms ,Epithelium ,Pathology and Forensic Medicine ,Cell biology ,Adult Stem Cells ,medicine.anatomical_structure ,Gene Expression Regulation ,Cancer stem cell ,Amniotic epithelial cells ,Immunology ,medicine ,Animals ,Humans ,Regeneration ,Respiratory epithelium ,Progenitor cell ,Stem cell ,Adult stem cell - Abstract
Epithelial branching during the process of lung development results in the establishment of distinct functional zones, each of which is characterized by a unique cellular composition and repertoire of local progenitor cells. Significant new insights into cellular and molecular mechanisms of epithelial maintenance that provide insights into the pathophysiology of lung disease have been made in recent years. This review focuses on the complex structure-function relationship in the airway epithelium, how this epithelium is maintained in the normal state and repaired following injury, and how deregulation may contribute to airway disease and cancer.
- Published
- 2009
49. Lgr4 and Lgr5 drive the formation of long actin-rich cytoneme-like membrane protrusions
- Author
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H. Kim Lyerly, Lauren K. Rochelle, Larry S. Barak, Marc G. Caron, Sébastien Marion, and Joshua C. Snyder
- Subjects
Adult ,Cell signaling ,Arrestins ,Blotting, Western ,Cell Surface Extension ,Biology ,Receptors, G-Protein-Coupled ,Cell membrane ,Cancer stem cell ,Myosin ,medicine ,Humans ,Immunoprecipitation ,Pseudopodia ,Molecular Biology ,Actin ,Tissue homeostasis ,beta-Arrestins ,Beta-Arrestins ,Stem Cells ,Cell Membrane ,LGR5 ,Biological Transport ,Cell Biology ,beta-Arrestin 2 ,Actins ,Cell biology ,medicine.anatomical_structure ,HEK293 Cells ,Cell Surface Extensions ,Filopodia ,Developmental Biology ,Cytoneme ,Signal Transduction ,Research Article - Abstract
Embryonic development and adult tissue homeostasis require precise information exchange between cells and their microenvironment to coordinate cell behavior. A specialized class of ultra-long actin-rich filopodia, termed cytonemes, provides one mechanism for this spatiotemporal regulation of extracellular cues. We provide here a mechanism whereby the stem-cell marker Lgr5, and its family member Lgr4, promote the formation of cytonemes. Lgr4- and Lgr5-induced cytonemes exceed lengths of 80 µm, are generated through stabilization of nascent filopodia from an underlying lamellipodial-like network and functionally provide a pipeline for the transit of signaling effectors. As proof-of-principle, we demonstrate that Lgr5-induced cytonemes act as conduits for cell signaling by demonstrating that the actin motor and filopodial cargo carrier protein myosin X (Myo10) and the G-protein-coupled receptor (GPCR) signaling effector β-arrestin-2 (Arrb2) transit into cytonemes. This work delineates a biological function for Lgr4 and Lgr5 and provides the rationale to fully investigate Lgr4 and Lgr5 function and cytonemes in mammalian stem cell and cancer stem cell behavior.
- Published
- 2015
50. Relevance of GPCR functional selectivity/biased signaling to drugs of abuse (468.1)
- Author
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Marc G. Caron, Nikhil M. Urs, Sean M. Peterson, Joshua C. Snyder, and Tanya L. Daigle
- Subjects
Drugs of abuse ,Cell signaling ,G protein ,Chemistry ,fungi ,food and beverages ,Pharmacology ,Biochemistry ,Genetics ,Functional selectivity ,Molecular Biology ,Neuroscience ,Intracellular ,Biotechnology ,G protein-coupled receptor - Abstract
We now understand that GPCRs can signal not only through activation of G proteins but also through the ability of β-arrestins to scaffold intracellular signaling molecules with distinct temporal an...
- Published
- 2014
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