Papers by Benjamin L . Bryson
Supplemental Information to: Aberrant induction of a mesenchymal/stem-cell program engages senesc... more Supplemental Information to: Aberrant induction of a mesenchymal/stem-cell program engages senescence in normal mammary epithelial cells S1. Cytokine-induced senescence induces EMT and SC-associated gene expression changes. S2. OSM-mediated Snail induction drives an aberrant EMT that induces senescence. S3. OSM-induced EMT requires STAT3- and TGF-β/SMAD3-mediated signaling. S4. Senescence-escape generates transformed CSC which are highly sensitive to palbociclib. S5. Proposed mechanisms of OSM induced senescence, EMT, and stemness in normal human epithelial cells versus transformed cells. S6. Uncropped images of all western blots presented in the current study (continued). Supplementary Table 1. List of primary antibodies used in this study Supplementary Table 2. qRT-PCR primer sequences (listed 5'-to-3') used in this study.
Journal of Biological Chemistry
Cancer Research
Mitotic kinases are essential for cell cycle progression, and several mitotic kinase inhibitors h... more Mitotic kinases are essential for cell cycle progression, and several mitotic kinase inhibitors have been developed to target the uncontrolled proliferation of cancer cells. However, despite their efficacy in preclinical models, mitotic kinase inhibitors are generally considered poor therapeutic targets due to their off target effects in other rapidly proliferating tissues and subsequent toxicity. However, preliminary data in our lab shows that relative to non-transformed cells, triple negative breast cancer (TNBC) cells are selectively sensitive to inhibition of the mitotic kinase NEK2 compared to other mitotic kinase inhibitors. Therefore, we hypothesize that NEK2 is a targetable vulnerability of TNBC. To begin to understand the role of NEK2 in TNBC, we performed RNA-sequencing following NEK2 silencing and found increased expression of genes important at the G1/S cell cycle transition. This transition is largely controlled by the RB-E2F signaling axis, where CDK4 and CDK6 are key ...
PDF file - 1239KB, S1. Expression of exogenous FAM83B relative to endogenous FAM83B. Western blot... more PDF file - 1239KB, S1. Expression of exogenous FAM83B relative to endogenous FAM83B. Western blot of control GFP-expressing HME1 cells and FAM83B-expressing HME1 cells using a FLAG antibody or a FAM83B antibody. S2. Expression of FAM83A, FAM83B and FAM83D in lung cancer subtypes. The samples included normal lung (Norm - 38 samples), squamous cell carcinoma (SCC - 30 samples), Adenocarcinoma (Ad - 33 samples), Adenosquamous carcinoma (AdS - 4 samples), non-small cell carcinoma (NSC - 9 samples), large cell carcinoma (LC - 12 samples), small cell carcinoma (SC - 3 samples). The significant p-values defined as follows, p<0.005 (*) and p<0.05 (#). S3. FAM83 member expression in normal tissues. Real-time PCR analysis was performed for FAM83 members A-E. The relative expression of each FAM83 member is presented from all normal, associated tissues from an Origene TissueScan Cancer Survey Panel. The expression of each FAM83 member in each tissue was normalized to its expression in the...
Supplementary information. (PDF 1290 kb)
Additional file 5: Table S1. ChIP-PCR primer sequences. Primer sequences targeting six regions wi... more Additional file 5: Table S1. ChIP-PCR primer sequences. Primer sequences targeting six regions within the EGFR promoter are listed.
Additional file 3: Figure S3. KLF4 negatively regulates the EGFR signaling pathway. a REVERT stai... more Additional file 3: Figure S3. KLF4 negatively regulates the EGFR signaling pathway. a REVERT staining of total protein in Fig. 3a. b REVERT staining of total protein in Fig. 3c. c REVERT staining of total protein in Fig. 3e.
Additional file 4: Figure S4. Repression of EGFR is an obligatory intermediate step for KLF4 to i... more Additional file 4: Figure S4. Repression of EGFR is an obligatory intermediate step for KLF4 to inhibit aggressive breast cancer phenotypes. a REVERT staining of total protein in Fig. 5a. b REVERT staining of total protein in Fig. 5b.
Additional file 1: Figure S1. KLF4 represses migration, invasion, and cell growth in breast epith... more Additional file 1: Figure S1. KLF4 represses migration, invasion, and cell growth in breast epithelial cells. a REVERT staining of total protein across eight different breast cell lines: MCF10A (10A), MCF7, T47D, SKBR3, SUM159, HCC70, MDA-MB-468 (468), and MDA-MB-231 (231). b REVERT staining of total protein in MCF10A cells as well as MDA-MB-231 and MDA-MB-468 after transduction with AdGFP (G) or AdKLF4 (K). c REVERT staining of total protein in MCF10A cells after transfection with siNS or siKLF4 (siK).
Additional file 2: Figure S2. Identification of a KLF4-regulated protein signature reveals EGFR a... more Additional file 2: Figure S2. Identification of a KLF4-regulated protein signature reveals EGFR as a downstream target. The top a 63 downregulated proteins and b 14 upregulated proteins after KLF4 overexpression. c Protein-Protein interaction network including proteins with greater than 2-fold expression decrease after AdKLF4 infection and RPPA analysis. Colored nodes indicate query proteins and the first shell of interactors. Teal and purple lines indicate known interactions. Green, red, yellow and blue lines indicate predicted interactions. d REVERT staining of total protein in MDA-MB-231 cells after transduction with AdGFP (AdG) or AdKLF4 (AdK).
FAM83B (Family with sequence similarity 83, member B) was recently identified as a novel oncogene... more FAM83B (Family with sequence similarity 83, member B) was recently identified as a novel oncogene involved in activating CRAF/MAPK signaling and driving epithelial cell transformation. FAM83B is one of eight members of a protein family (FAM83) characterized by a highly conserved domain of unknown function (DUF1669), which is necessary and sufficient to drive transformation. Here, it is demonstrated that additional FAM83 members also exhibit oncogenic properties and have significantly elevated levels of expression in multiple human tumor types using a TissueScan Cancer Survey Panel PCR array and database mining. Furthermore, modeling the observed tumor expression of FAM83A, FAM83C, FAM83D, or FAM83E promoted human mammary epithelial cell (HMEC) transformation, which correlated with the ability of each FAM83 member to bind CRAF (RAF1) and promote CRAF membrane localization. Conversely, ablation of FAM83A or FAM83D from breast cancer cells resulted in diminished MAPK signaling with marked suppression of growth in vitro and tumorigenicity in vivo. Importantly, each FAM83 member was determined to be elevated in at least one of 17 distinct tumor types examined, with FAM83A, FAM83B, and FAM83D most frequently overexpressed in several diverse tissue types. Finally, evidence suggests that elevated expression of FAM83 members is associated with tumor grade and overall survival. Implications: FAM83 proteins represent a novel family of oncogenes suitable for the development of cancer therapies aimed at suppressing MAPK signaling.
www.mdpi.com/journal/cancers
Molecular Cancer Research, 2020
Although frequently associated with tumor progression, inflammatory cytokines initially restrain ... more Although frequently associated with tumor progression, inflammatory cytokines initially restrain transformation by inducing senescence, a key tumor-suppressive barrier. Here, we demonstrate that the inflammatory cytokine, oncostatin M, activates a mesenchymal/stem cell (SC) program that engages cytokine-induced senescence (CIS) in normal human epithelial cells. CIS is driven by Snail induction and requires cooperation between STAT3 and the TGFβ effector, SMAD3. Importantly, as cells escape CIS, they retain the mesenchymal/SC program and are thereby bestowed with a set of cancer SC (CSC) traits. Of therapeutic importance, cells that escape CIS can be induced back into senescence by CDK4/6 inhibition, confirming that the mechanisms allowing cells to escape senescence are targetable and reversible. Moreover, by combining CDK4/6 inhibition with a senolytic therapy, mesenchymal/CSCs can be efficiently killed. Our studies provide insight into how the CIS barriers that prevent tumorigenesi...
Journal of Biological Chemistry, 2021
Breast Cancer Research, 2020
Background Triple-negative breast cancer (TNBC) is characterized by high rates of recurrence and ... more Background Triple-negative breast cancer (TNBC) is characterized by high rates of recurrence and poor overall survival. This is due, in part, to a deficiency of targeted therapies, making it essential to identify therapeutically targetable driver pathways of this disease. While epidermal growth factor receptor (EGFR) is expressed in 60% of TNBCs and drives disease progression, attempts to inhibit EGFR in unselected TNBC patients have had a marginal impact on outcomes. Hence, we sought to identify the mechanisms that dictate EGFR expression and inhibitor response to provide a path for improving the utility of these drugs. In this regard, the majority of TNBCs express low levels of the transcription factor, Krüppel-like factor 4 (KLF4), while a small subset is associated with high expression. KLF4 and EGFR have also been reported to have opposing actions in TNBC. Thus, we tested whether KLF4 controls the expression of EGFR and cellular response to its pharmacological inhibition. Metho...
Oncogene, 2019
The serine/threonine Protein Phosphatase 2A (PP2A) functions as a tumor suppressor by negatively ... more The serine/threonine Protein Phosphatase 2A (PP2A) functions as a tumor suppressor by negatively regulating multiple oncogenic signaling pathways. The canonical PP2A holoenzyme is comprised of a scaffolding subunit (PP2A Aα/β), which serves as the platform for binding of both the catalytic C subunit and one regulatory B subunit. Somatic heterozygous missense mutations in PPP2R1A, the gene encoding the PP2A Aα scaffolding subunit, have been identified across multiple cancer types, but the effects of the most commonly mutated residue, Arg-183, on PP2A function have yet to be fully elucidated. In this study, we used a series of cellular and in vivo models and discovered that the most frequent Aα R183W mutation formed alternative holoenzymes by binding of different PP2A regulatory subunits compared to wild type Aα, Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:
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Papers by Benjamin L . Bryson