The Greenfield Lab (Indianapolis) has long-standing research interests focusing on basic and translational aspects of bone biology, and on osteosarcoma metastasis.
Greenfield Lab
The Greenfield Lab’s long-standing research interests focus on basic and translational aspects of bone biology, and on osteosarcoma metastasis.
The osteosarcoma project aims to repurpose FDA-approved drugs to block growth of lung metastases, which is the primary cause of lethality in osteosarcoma patients. Current studies utilize 3D sarcosphere cultures and mouse models.
The bone biology projects aim to understand how aging, inflammation, infection, and PKA signaling regulate bone turnover, fracture healing, and implant integration/loosening. Current studies utilize cell cultures (osteoblasts, osteoclasts, and macrophages) and murine models (transgenic and surgical).
Principal Investigator
Ed Greenfield, PhD
Professor of Orthopaedic Surgery
Ed Greenfield, PhD, joined the IU School of Medicine Department of Orthopaedic Surgery in 2019 after a long career on the faculty at Case Western Reserve University in Cleveland. His affiliations, in addition to the Department of Orthopaedic Surgery, include the Indiana Center for Musculoskeletal Health and the IU Health Simon Cancer Center.
To stay current on the medical research work at IU School of Medicine, follow the IU School of Medicine research blog, where investigators throughout the school’s academic departments statewide post updates about their work.
The major goals of the Greenfield Lab’s osteosarcoma project are to:
1. Determine which HDAC inhibitors and proteasome inhibitors are most promising for osteosarcoma patients, and
2. Determine whether combinations of HDAC inhibitors, proteasome inhibitors, and/or standard-of-care MAP chemotherapeutics are more promising than monotherapy.
The major goal of the Greenfield Lab’s aging project is to determine whether PKI-gamma mediates age related bone loss and the age related decline in skeletal healing by regulating the balance between osteogenesis and adipogenesis in a sex- and skeletal site-dependent manner.
The major goals of the Greenfield Lab’s implant loosening project are:
1. To determine whether experimentally-induced periodontitis increases particle-induced osteolysis in mice via systemic translocation of PAMPs or bacteria, and
2. To determine whether PAMPs, periodontal disease, nasal colonization, or the bacterial biofilms on failed joint replacements correlate with peri-prosthetic inflammation or particle-induced osteolysis in human patients without clinical signs of infection.
The major goal of Dr. Greenfield’s implant infection project is to determine whether Halicin, a novel anti-bacterial agent, is effective against bacterial biofilms in vitro and in mice.
Important Findings
HDAC inhibitors, proteasome inhibitors, and tyrosine kinase inhibitors block growth of 3D osteosarcoma spheroids.
PKI-gamma mediates age related bone loss and the age related decline in skeletal healing by regulating cAMP/PKA signaling.
Inflammation and osteolysis induced by orthopaedic wear particles are increased by adherent bacterial PAMPs.
Lab Staff
The Greenfield Lab is powered by skilled postdoctoral and graduate-level researchers, lab techs and managers.
Opportunities are available to learners interested in research.
Residents and medical students can join projects that focus on repurposing FDA-approved drugs to block growth of osteosarcoma metastases or on how aging, inflammation, infection, and PKA signaling regulate bone turnover, fracture healing, and implant integration/loosening. They would learn in vivo mouse models and/or cell culture models (3D sarcospheres, osteoblasts, osteoclasts, and macrophages) and murine models (transgenic and surgical).
Current Staff Members
Shota Higashihira, MD, PhD, Postdoctoral Research Fellow
Emily Seiden, BS, Graduate Student Researcher
Stefanie Simpson, MS, Lab Manager
Piper Wilburn, BS, Lab Tech
Publications
Fort BP, Dubyak GR, Greenfield EM. Lysosomal disruption by orthopedic wear particles induces activation of the NLRP3 inflammasome and macrophage cell death by distinct mechanisms. J Orthop Res. 2021 Mar;39(3):493-505. doi: 10.1002/jor.24826. Epub 2020 Aug 22. PMID: 32779803; PMCID: PMC8201664.
Collier CD, Hausman BS, Zulqadar SH, Din ES, Anderson JM, Akkus O, Greenfield EM. Characterization of a reproducible model of fracture healing in mice using an open femoral osteotomy. Bone Rep. 2020 Feb 5;12:100250. doi: 10.1016/j.bonr.2020.100250. Erratum in: Bone Rep. 2021 Apr 29;14:101084. PMID: 32090156; PMCID: PMC7025178.
Collier CD, Getty PJ, Greenfield EM. Targeting the Cancer Epigenome with Histone Deacetylase Inhibitors in Osteosarcoma. Adv Exp Med Biol. 2020;1258:55-75. doi: 10.1007/978-3-030-43085-6_4. PMID: 32767234.
Manzano GW, Fort BP, Dubyak GR, Greenfield EM. Wear Particle-induced Priming of the NLRP3 Inflammasome Depends on Adherent Pathogen-associated Molecular Patterns and Their Cognate Toll-like Receptors: An In Vitro Study. Clin Orthop Relat Res. 2018 Dec;476(12):2442-2453. doi: 10.1097/CORR.0000000000000548. PMID: 30427314; PMCID: PMC6259896.
Bechtel CP, Gebhart JJ, Tatro JM, Kiss-Toth E, Wilkinson JM, Greenfield EM. Particle-Induced Osteolysis Is Mediated by TIRAP/Mal in Vitro and in Vivo: Dependence on Adherent Pathogen-Associated Molecular Patterns. J Bone Joint Surg Am. 2016 Feb 17;98(4):285-94. doi: 10.2106/JBJS.O.00736. PMID: 26888676.
Choe H, Narayanan AS, Gandhi DA, Weinberg A, Marcus RE, Lee Z, Bonomo RA, Greenfield EM. Immunomodulatory Peptide IDR-1018 Decreases Implant Infection and Preserves Osseointegration. Clin Orthop Relat Res. 2015 Sep;473(9):2898-907. doi: 10.1007/s11999-015-4301-2. PMID: 25953690; PMCID: PMC4523515.
Bonsignore LA, Goldberg VM, Greenfield EM. Machine oil inhibits the osseointegration of orthopaedic implants by impairing osteoblast attachment and spreading. J Orthop Res. 2015 Jul;33(7):979-87. doi: 10.1002/jor.22850. Epub 2015 May 21. PMID: 25676177; PMCID: PMC8201705.
Chen X, Hausman BS, Luo G, Zhou G, Murakami S, Rubin J, Greenfield EM. Protein kinase inhibitor γ reciprocally regulates osteoblast and adipocyte differentiation by downregulating leukemia inhibitory factor. Stem Cells. 2013 Dec;31(12):2789-99. doi: 10.1002/stem.1524. PMID: 23963683; PMCID: PMC3930625.
Bonsignore LA, Anderson JR, Lee Z, Goldberg VM, Greenfield EM. Adherent lipopolysaccharide inhibits the osseointegration of orthopedic implants by impairing osteoblast differentiation. Bone. 2013 Jan;52(1):93-101. doi: 10.1016/j.bone.2012.09.011. Epub 2012 Sep 17. PMID: 22995462; PMCID: PMC3513552.
Rettew AN, Young ED, Lev DC, Kleinerman ES, Abdul-Karim FW, Getty PJ, Greenfield EM. Multiple receptor tyrosine kinases promote the in vitro phenotype of metastatic human osteosarcoma cell lines. Oncogenesis. 2012 Nov 19;1(11):e34. doi: 10.1038/oncsis.2012.34. PMID: 23552467; PMCID: PMC3511679.
Funding
Current funding sources include:
2020-2025
PI (29% effort). NIH/NIA R01. Regulation of age-related bone loss by PKI-gamma. TDC: $1,305,629/ 5 years.
2020-2021
Multi-PI with Erik Imel & Yasuyoshi Ueki, Indiana Center for Musculoskeletal Health multi-PI Pilot Award, Bacterial contribution to wear particle-induced failure of orthopaedic joint replacement prostheses in patients without clinical signs of infection, TDC $40,000/ 1 year
2021-2022
Multi-PI with Karen Pollok, IU Health Simon Cancer Center EDT/TMM Pilot Award (no salary), Repurposing HDAC inhibitors and proteasome inhibitors to increase survival in pre-clinical models of established osteosarcoma metastases, TDC $49,998/ 1 year
2021-2022
PI (no salary), Indiana CTSI Pilot Funding for Research Use of Core Facilities, Regulation of Macrophage Subsets by Orthopaedic Wear Particles, TDC $10,000/ 1 year
