Michael Goldstein Lab
Our lab is studying the role of epigenetic pathways in cellular DNA damage responses with the main goal to identify new molecular targets in tumors and improve cancer care.
We investigate the molecular mechanisms of cellular responses to radiation- and chemotherapy-induced DNA damage, specifically focusing on glioblastoma, medulloblastoma and cervical cancer as tumor models. Our primary aim is to identify new DNA damage response pathways that can be pharmacologically targeted in order to improve tumor response to genotoxic treatment by radiation and chemotherapy. We are specifically interested in epigenetic changes that occur at the sites of DNA damage including alteration of chromatin structure and post-translational modifications of histone proteins. These events are critical for repair of DNA lesions making epigenetic modifiers attractive targets for sensitization of cancer cells to radiotherapy and chemotherapy.
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DDR pathways involved in tumor response to anti-cancer agents
From Goldstein M and Kastan B, Annual Reviews Medicine, 2015 (source)
Responses to DNA damage are important determinants of cell viability and mutagenesis. Following DNA damage induction, an intricate network of DDR signaling pathways is activated to enforce cell cycle arrest, DNA repair, and potentially cell death. The ability of a cell to detect and repair damaged DNA has a direct effect on cell survival after exposure to genotoxic stress. Importantly, many agents that are clinically used to treat cancer function by damaging DNA. Therefore, the ability of tumor cells to cope with DNA damage induced by radiation and chemotherapy affects the efficacy of these treatment modalities and disease outcomes. DNA double-strand breaks (DSBs) are the most toxic DNA lesions and are considered the major trigger of cell death induced by radiation and chemotherapy. Consequently, a specific inhibition of DSB repair in cancer cells can improve radiation response and the overall outcome in cancer patients.
Epigenetic alteration of chromatin including chromatin structure changes and post-translational modifications (PTMs) of histone proteins collectively form a complex epigenetic landscape regulating transcription, replication, and DDR. We have discovered that eviction of the H2A/H2B histone dimers is facilitated by nucleolin at the DSB sites, an event that is critical for repair of the DNA breaks by non-homologous end-joining. Additionally, multiple histone PTMs have been identified that promote recruitment of DDR factors to the break. For instance, a simultaneous methylation of H4K20 and ubiquitination of H2AK15 is required for 53BP1 recruitment and, consequently, DSB repair demonstrating how an intricate histone PTM network facilitates DDR.
Acetylation of histone lysine residues occurs frequently throughout the entire epigenome and plays an important role in transcription and DDR. A recent discovery of novel short-chain histone lysine acylations including propionylation, crotonylation succinylation and β-hydroxy-butyrylation has extended the arsenal of histone modifications. Increasing evidence suggests that these PTMs regulate cellular processes such as transcriptional activity. We are studying the role of these novel histone modifications in cellular response to radiation-induced DSBs. Our goal is to identify specific histone sites that are modified by these acylations at the DSB. Further, we are interested in dissecting the molecular mechanisms through which these modifications regulate DDR. We are employing multiple experimental techniques including laser-microirradiation, immuno-fluorescence, endonuclease-based DNA repair assays and chromatin immunoprecipitation in order to elucidate the role of these histone PTMs in DDR.
Glioblastoma is one of the most frequent and the most aggressive primary malignant brain affecting both, pediatric and adult patients. Despite some advances in treatment, prognosis of glioblastoma remains poor with less than 5% of patients surviving 5 years after diagnosis. An adjuvant temozolomide chemotherapy combined with radiotherapy represents the standard-of-care for glioblastoma. Thus, tumor response to radiation and temozolomide is critical for patient survival. Temozolomide is an alkylating agent that induces methylation of DNA bases. Whereas, radiation directly induces DSBs, temozolomide-induced DNA lesions have to be converted into DSBs in a mismatch repair and proliferation dependent manner.
Glioblastoma is characterized by a frequent deregulation of epigenetic pathways due to mutations of histone modifiers. We are investigating epigenetic pathways that when defective result in glioblastoma resistance to the standard-of-care chemoradiotherapy. Furthermore, we are developing strategies to restore chemoradiotherapy response in these tumors by using targeted therapeutics. This investigation will contribute to the personalized medicine approach in treatment of glioblastoma by tailoring treatment based on the individual mutation profile of epigenetic modifiers and has a potential to improve outcomes of this aggressive disease.
Radiation therapy is an important treatment modality of locally advanced cervical cancer. Thus, understanding the mechanisms of radiation response in cervical cancer is critical for development of new treatment strategies that can overcome radiation resistance and improve survival in patients. Radiation kills cancer cells by inducing toxic DNA doubles-strand breaks (DSBs). Increasing evidence suggests that epigenetic changes at the sites of radiation-induced DSBs play a critical role in DNA repair. Identifying epigenetic pathways that are involved in radiation response in cervical cancer cells can result in development of new therapeutics that can overcome radiation resistance and improve the outcomes of the disease.
Infection with high-risk HPV subtypes, including HPV 16 and 18, is a common driver of cervical carcinogenesis, associated with over 80% of cervical cancers. The oncogenic properties of the HPV genome have been attributed to the E6 and E7 gene products that affect multiple cellular processes including cell cycle, DNA repair and epigenetic pathways such as histone acetylation and methylation. Consequently, expression of high-risk E6 and E7 genes results in an altered epigenetic landscape in HPV+ cancers.
We are using CRISPR/Cas9-based screening techniques to identify epigenetic modifiers that are involved in radiation response in HPV+ and HPV- cervical cancer. Further, we are elucidating the molecular mechanisms through which these epigenetic modifiers regulate DDR. Our ultimate goal is to identify new targets for radiosensitization that are specific for HPV+ and HPV- tumors.
Proton radiotherapy is an emerging treatment for medulloblastoma due to its ability to restrict radiation doses to the tumor, while sparing healthy tissues. Whereas proton radiation also kills tumor cells by generating DSBs, proton-induced DNA breaks differ from those induced by conventional γ-radiation (photon radiation). A characteristic of proton radiation is its high linear energy transfer (LET) in irradiated tissues, resulting in highly complex, clustered DNA lesions that persist for a longer period of time. Clustered DSBs require orchestration of multiple DNA processing and repair pathways, including NHEJ, HR, and base excision repair (BER) for repair and survival of proton- induced DNA damage.
We are studying the differences in molecular responses to DNA damage induced by photon versus proton radiation. These investigations will extend our knowledge of radiation biology and identify potential targets for sensitization of tumors to different types of radiotherapy.
Dr. Goldstein received his MD and PhD degrees from the Johannes Gutenberg University located in Mainz, Germany in 2008. As a graduate student he investigated the molecular mechanisms of the toxicity of chemotherapeutic agents used to treat cancer under the direction of Dr. Bernd Kaina.
He went on to complete a postdoctoral fellowship at St. Jude Children’s Research Hospital and Duke University in Dr. Michael Kastan’s lab. During this time his research focused on regulation of DNA double-strand break repair by modulation of chromatin structure at the DNA damage sites.
After finishing the postdoctoral fellowship in 2015 Dr. Goldstein completed an Internship in Internal Medicine at Duke University Medical Center. In 2016 he joined the Department of Radiation Oncology at the Washington University in St. Louis to establish his laboratory and is currently continuing his training as a resident in Radiation Oncology.
Dr. Goldstein’s current research focuses on the role of epigenetic pathways in DNA damage responses that tumor cells activate following treatment with radiation and chemotherapy. His lab performs both, basic and translational research with the ultimate goal of increasing the efficacy of radiotherapy and chemotherapy and improving cancer patient care.
In his free time he enjoys spending time in the gym, running, hiking, cooking dishes from all over the world and watching surreal movies.
Dr. Nishanth Gabriel joined the Goldstein lab in March 2017 as a Postdoctoral Research Associate. He is investigating the role of HPV infection in epigenetic regulation of DNA damage response in cervical cancers. He is also studying the role of histone modifiers in radiation and chemotherapy response in glioblastoma. He received his master’s degree in Biotechnology from Cochin University of Science and Technology, Cochin, Kerala, India. His Master’s thesis was to evaluate a protocol for easier HPV screening from cervical samples received from remote places without laboratory access. He pursued his doctoral research in Christian Medical College, Vellore, India where he elucidated the role of the Notch signaling pathway in human bone marrow derived stromal cells. He has also an extensive research experience with adenoviral vectors and gene therapy. Outside the lab, he enjoys music and tries to understand the art of investing.
Kumaresh Balaji is currently a post-baccalaureate pre-med student at University College, as part of Washington University in St. Louis. He graduated from Villanova University in 2015 with a B.S. in Cell and Molecular Biology. After graduating, he worked as a Biology Intern at WAVE Life Sciences, a medical genetics company, where his work focused on stereopure oligonucleotide targeting of genes and mutations underlying neuromuscular diseases. He plans on beginning medical school after completing his post-baccalaureate program.
In his free time Kumaresh enjoys visiting art museums and exhibits, reading creative nonfiction, and playing basketball.
Mallory Moran, BS (Post-baccalaureate student)
Mallory Moran is a student in the Post-Baccalaureate Premedical program at Washington University in St. Louis. She received her B.S. in Psychology from Pennsylvania State University, and plans to continue her education in medical laboratory science.
Mallory enjoys running, traveling as often as possible, and spending time with friends and family.
Wang Q, Goldstein M. Small RNAs recruit chromatin modifying enzymes MMSET and Tip60 to reconfigure damaged DNA upon double-strand break and facilitate repair. Cancer Res. 2016 Jan 28. pii: canres.2334.2015
Goldstein M, Kastan MB. Repair versus checkpoint functions of Brca1 are differentially regulated by site of chromatin binding. Cancer Res. 2015 Jul 1;75(13):2699-707
Goldstein M, Kastan M. The DNA Damage Response: Implications for Tumor Responses to Radiation and Chemotherapy. Ann Rev Med. 2015 Jan 14;66:129-43. doi: 10.1146/annurev-med-081313-121208.
Henriksson S, Rassoolzadeh H, Hedström E, Coucoravas C, Julner A, Goldstein M, Imreh G, Zhivotovsky B, Kastan MB, Helleday T, Farnebo M. The scaffold protein WRAP53β orchestrates the ubiquitin response critical for DNA double-strand break repair. Genes & Development. 2014 Dec 15;28(24):2726-38. doi: 10.1101/gad.246546.114.
Kennedy EM, Kornepati AV, Goldstein M, Bogerd HP, Poling BC, Whisnant AW, Kastan MB, Cullen BR. Inactivation of the human papillomavirus E6 or E7 gene in cervical carcinoma cells using a bacterial CRISPR/Cas RNA-guided endonuclease. J Virol. 2014 Oct; 88(20):11965-72. doi: 10.1128/JVI.01879-14. Epub 2014 Aug 6.2014
Wang Q, Goldstein M, Alexander P, Wakeman TP, Sun T, Feng J, Lou Z, Kastan MB, Wang XF. Rad17 recruits the MRE11-RAD50-NBS1 complex to regulate the cellular response to DNA double-strand breaks. EMBO J. 2014 Apr 16;33(8):862-77. Epub 2014 Feb 16.
Goldstein M, Derheimer FA, Tait-Mulder J, Kastan MB. Nucleolin Mediates Nucleosome Disruption Critical for DNA Double-Strand Break Repair. Proc Natl Acad Sci U S A. 2013 Oct 15; 110(42):16874-9. doi: 10.1073/pnas.1306160110. Epub 2013 Sep 30
Bauer M*, Goldstein M*, Heylmann D, Kaina B. Human monocytes undergo excessive apoptosis following temozolomide activating the ATM/ATR pathway while dendritic cells and macrophages are resistant. PLoS One. 2012; 7(6):e39956. Epub 2012 Jun 29.
Bauer M, Goldstein M, Christmann M, Becker H, Heylmann D, Kaina B. Human monocytes are severely impaired in base and DNA double-strand break repair that renders them vulnerable to oxidative stress. Proc Natl Acad Sci U S A. 2011 Dec 27; 108(52):21105-10. Epub 2011 Dec 12.
Goldstein M, Roos WP, Kaina B. Apoptotic death induced by the cyclophosphamide analogue mafosfamide in human lymphoblastoid cells: contribution of DNA replication, transcription inhibition and Chk/p53 signaling. Toxicol Appl Pharmacol. 2008 May 15; 229(1):20-32, 2008. Epub 2008 Jan 17