A5015351867- DNA Repair Mechanisms
- CRISPR and Genetic Engineering
- Advanced biosensing and bioanalysis techniques
- Cancer Genomics and Diagnostics
- PARP inhibition in cancer therapy
- Cancer therapeutics and mechanisms
- Epigenetics and DNA Methylation
- Genomics and Chromatin Dynamics
- Carcinogens and Genotoxicity Assessment
- Microtubule and mitosis dynamics
- Histone Deacetylase Inhibitors Research
- Advanced Fluorescence Microscopy Techniques
- Effects of Radiation Exposure
- Ubiquitin and proteasome pathways
- Peptidase Inhibition and Analysis
- Tannin, Tannase and Anticancer Activities
- Polyomavirus and related diseases
- Connexins and lens biology
- Neuroendocrine Tumor Research Advances
- PI3K/AKT/mTOR signaling in cancer
- Cell Adhesion Molecules Research
- DNA and Nucleic Acid Chemistry
- Tea Polyphenols and Effects
- Metal complexes synthesis and properties
- Angiogenesis and VEGF in Cancer
Michigan State University
2022-2025
Wayne State University
2015-2024
The Barbara Ann Karmanos Cancer Institute
2016-2023
Nonhomologous end-joining (NHEJ) and homologous recombination (HR) are the primary pathways for repairing DNA double-strand breaks (DSBs) during interphase, whereas microhomology-mediated (MMEJ) has been regarded as a backup mechanism. Through CRISPR-Cas9-based synthetic lethal screens in cancer cells, we identified subunits of 9-1-1 complex (RAD9A-RAD1-HUS1) its interacting partner, RHINO, crucial MMEJ factors. We uncovered an unexpected function RHINO restricting to mitosis. accumulates M...
ERCC1-XPF heterodimer is a 5'-3' structure-specific endonuclease which essential in multiple DNA repair pathways mammalian cells. (ERCC1-ERCC4) repairs cisplatin-DNA intrastrand adducts and interstrand crosslinks its specific inhibition has been shown to enhance cisplatin cytotoxicity cancer In this study, we describe high throughput screen (HTS) used identify small molecules that inhibit the activity of ERCC1-XPF. Primary screens identified two compounds nanomolar range. These were...
Abstract Ubiquitin-specific peptidase 10 (USP10) stabilizes both tumor suppressors and oncogenes in a context-dependent manner. However, the nature of USP10’s role non-small cell lung cancer (NSCLC) remains unclear. By analyzing The Cancer Genome Atlas (TCGA) database, we have shown that high levels USP10 are associated with poor overall survival NSCLC mutant p53, but not wild-type p53. Consistently, genetic depletion or pharmacological inhibition dramatically reduces growth xenografts...
Repair of DNA double strand breaks (DSBs) is integral to preserving genomic integrity. Therefore, defining the mechanisms underlying DSB repair will enhance our understanding how defects in these pathways contribute human disease and could lead discovery new approaches for therapeutic intervention. Here, we established a panel HaloTagged damage response factors U2OS cells which enables concentration-dependent protein labeling by fluorescent HaloTag ligands. Genomic insertion at endogenous...
DNA double strand breaks (DSBs) are widely considered the most cytotoxic lesions occurring in cells because they physically disrupt connectivity of helix. Homologous recombination (HR) is a high-fidelity DSB repair pathway that copies sequence spanning break from homologous template, commonly sister chromatid. How both ends, and chromatid held close proximity during HR unknown. Here we demonstrate PST repeat region MDC1 mutlivalent nucleosome binding domain, sufficient to tether chromatin...
Protein tagging with CRISPR-Cas9 enables the investigation of protein function in its native environment but is limited by low homology-directed repair (HDR) efficiency causing knock-in rates. We present a detailed pipeline using HDR donor plasmids containing antibiotic resistance cassettes for rapid selection gene-edited cells. Our protocol streamlines N- or C-terminal human cells, enabling plasmid preparation single cloning step.
The 5'-3' structure-specific endonuclease ERCC1/XPF (Excision Repair Cross-Complementation Group 1/Xeroderma Pigmentosum group F) plays critical roles in the repair of cisplatin-induced DNA damage. As such, it has been identified as a potential pharmacological target for enhancing clinical response to platinum-based chemotherapy. goal this study was follow up on our previous identification compound NSC143099 potent inhibitor activity by performing an silico screen identify structural...
The radiation-induced bystander effect (RIBE) can increase cellular toxicity in a gap junction dependent manner unirradiated cells. Recent reports have suggested that cisplatin also be mediated by functional intercellular communication (GJIC). In this study using lung and ovarian cancer cell lines, we showed cytotoxicity is density. This ablated when GJA1 or Connexin 43 (Cx43) targeted, gene protein, respectively, leading to resistance but only at high forming We observed the...
Abstract Purpose: ERCC1/XPF is a DNA endonuclease with variable expression in primary tumor specimens, and has been investigated as predictive biomarker for efficacy of platinum-based chemotherapy. The failure clinical trials utilizing ERCC1 to predict response chemotherapy suggests additional mechanisms underlying the basic biology interstrand crosslinks (ICLs) remain unknown. We aimed characterize panel knockout (Δ) cell lines, where we identified synthetic viable phenotype ICLs...
DNA double-strand breaks (DSBs) are toxic lesions that can lead to genome instability if not properly repaired. Breaks incurred in G1 phase of the cell cycle predominantly fixed by non-homologous end-joining (NHEJ), while homologous recombination (HR) is primary repair pathway S and G2. Microhomology-mediated (MMEJ) intrinsically error-prone considered a backup DSB becomes essential when HR NHEJ compromised. In this study, we uncover MMEJ as major M phase. Using CRISPR/Cas9-based synthetic...
Carcinoma-associated fibroblasts (CAFs), a prominent cell type in the tumor microenvironment (TME), significantly contributes to cancer progression through interactions with cells and other TME components. Consequently, targeting signaling pathways driven by CAFs has potential yield new therapeutic approaches inhibit progression. However, mechanisms underlying their long-term vivo remains poorly understood.
Non-homologous end joining (NHEJ) is the predominant pathway that repairs DNA double-stranded breaks (DSBs) in vertebrates. However, due to challenges detecting DSBs living cells, repair capacity of NHEJ unknown. The termini many must be processed allow ligation while minimizing genetic changes result from break repair. Emerging models propose are first synapsed ~115 Å apart one several long-range synaptic complexes before transitioning into a short-range complex juxtaposes ends facilitate...
Replication protein A (RPA) plays essential roles in DNA replication, repair, recombination, and the damage response (DDR). Retrospective analysis of lung cancer patient data demonstrates high RPA expression as a negative prognostic biomarker for overall survival smoking-related cancers. Similarly, relative is predictive marker to chemotherapy. These observations are consistent with increase serving an adaptive mechanism that allows tolerance genotoxic stress resulting from carcinogen...
SUMMARY Non-homologous end joining (NHEJ) is the predominant pathway that repairs DNA double-stranded breaks (DSBs) in vertebrates. However, due to challenges detecting DSBs living cells, repair capacity of NHEJ unknown. The termini many must be processed allow ligation while minimizing genetic changes result from break repair. Emerging models propose are first synapsed ~115Å apart one several long-range synaptic complexes before transitioning into a short-range complex juxtaposes ends...
Platinum-based chemotherapy remains a mainstay treatment for the management of advanced non-small cell lung cancer. A key cellular factor that contributes to sensitivity platinums is 5′-3′ structure-specific endonuclease excision repair cross-complementation group 1 (ERCC1)/ xeroderma pigmentosum F (XPF). ERCC1/XPF critical platinum-induced DNA damage and has been subject intense research efforts identify small molecule inhibitors its nuclease activity purpose enhancing patient response...
Abstract ERCC1/XPF is a heterodimeric DNA endonuclease critical for repair of certain chemotherapeutic agents. We recently identified that ERCC1- and p53-deficient lung cancer cells are tolerant to platinum-based chemotherapy. ATR inhibition synergistically re-stored platinum sensitivity ERCC1-deficient cells. Mechanistically we show this effect reliant upon several functions including replication fork protection altered cell cycle checkpoints. Utilizing an inhibitor protein A (RPA), further...
Uncontrolled degradation and collapse of stalled replication forks (RFs) are primary sources genomic instability, yet the molecular mechanisms for protecting from degradation/collapse remain to be fully elaborated. Here, we show that polynucleotide kinase-phosphatase (PNKP) localizes at protects excessive degradation. The loss PNKP results in nucleolytic nascent DNA RFs. This mechanism is different BRCA2-dependent fork protection pathway, which MRE11-dependent Our research shows hydroxyurea...
ABSTRACT Repair of DNA double strand breaks (DSBs) is integral to preserving genomic integrity. Therefore, defining the mechanisms underlying DSB repair will enhance our understanding how defects in these pathways contribute human disease and could lead discovery new approaches for therapeutic intervention. Here, we established a panel HaloTagged damage response factors U2OS cells which enables concentration-dependent protein labeling. Genomic insertion HaloTag at endogenous loci preserves...
Abstract Replication protein A (RPA) plays essential roles in DNA replication, repair, recombination and the DNA-damage response (DDR). We have developed second generation RPA inhibitors (RPAi’s) that block RPA-DNA interaction. These DNA-binding (DBi’s) can elicit a state of cellular exhaustion resulting single agent vitro anticancer activity across broad spectrum cancers vivo two non-small cell lung cancer models. The to RPAi treatment suggests threshold exists before inhibition induces...
<p>Supplemental Figure S1. A. Clonogenic survival assays with cisplatin treatment in p53WT cell lines {plus minus} ERCC1. B. p53 null and p53mutant n = 3 independent experiments plated triplicate for each line. Data plotted as average of at least three minus}SD. C. Viability D. p53null SD. Supplemental S2. Cell utilized the current study status p53, EGFR, K-ras are listed. Mutation was obtained from Cosmic Database (cancer.sanger.ac.uk) or cBIOportal (cbioportal.org) (except H522 where...
<p>Supplemental Figure S1. A. Clonogenic survival assays with cisplatin treatment in p53WT cell lines {plus minus} ERCC1. B. p53 null and p53mutant n = 3 independent experiments plated triplicate for each line. Data plotted as average of at least three minus}SD. C. Viability D. p53null SD. Supplemental S2. Cell utilized the current study status p53, EGFR, K-ras are listed. Mutation was obtained from Cosmic Database (cancer.sanger.ac.uk) or cBIOportal (cbioportal.org) (except H522 where...
<div>AbstractPurpose:<p>ERCC1/XPF is a DNA endonuclease with variable expression in primary tumor specimens, and has been investigated as predictive biomarker for efficacy of platinum-based chemotherapy. The failure clinical trials utilizing ERCC1 to predict response chemotherapy suggests additional mechanisms underlying the basic biology interstrand crosslinks (ICLs) remain unknown. We aimed characterize panel knockout (Δ) cell lines, where we identified synthetic viable...