Visualizing Engrafted Human Cancer and Therapy Responses in Immunodeficient Zebrafish

Chuan Yan; Dalton C Brunson; Qin Tang; Daniel Do; Nicolae A Iftimia; John C Moore; Madeline N Hayes; Alessandra M Welker; Elaine G Garcia; Taronish D Dubash; Xin Hong; Benjamin J Drapkin; David T Myers; Sarah Phat; Angela Volorio; Dieuwke L Marvin; Matteo Ligorio; Lyle Dershowitz; Karin M McCarthy; Murat N Karabacak; Jonathan A Fletcher; Dennis C Sgroi; John A Iafrate; Shyamala Maheswaran; Nick J Dyson; Daniel A Haber; John F Rawls; David M Langenau

doi:10.1016/j.cell.2019.04.004

Visualizing Engrafted Human Cancer and Therapy Responses in Immunodeficient Zebrafish

Cell. 2019 Jun 13;177(7):1903-1914.e14. doi: 10.1016/j.cell.2019.04.004. Epub 2019 Apr 25.

Authors

Chuan Yan¹, Dalton C Brunson², Qin Tang², Daniel Do², Nicolae A Iftimia¹, John C Moore¹, Madeline N Hayes¹, Alessandra M Welker¹, Elaine G Garcia¹, Taronish D Dubash³, Xin Hong³, Benjamin J Drapkin³, David T Myers³, Sarah Phat³, Angela Volorio⁴, Dieuwke L Marvin³, Matteo Ligorio³, Lyle Dershowitz¹, Karin M McCarthy¹, Murat N Karabacak⁵, Jonathan A Fletcher⁶, Dennis C Sgroi⁴, John A Iafrate⁴, Shyamala Maheswaran³, Nick J Dyson³, Daniel A Haber⁷, John F Rawls⁸, David M Langenau⁹

Affiliations

¹ Molecular Pathology Unit, Massachusetts General Hospital Research Institute, Charlestown, MA 02129, USA; Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02139, USA.
² Molecular Pathology Unit, Massachusetts General Hospital Research Institute, Charlestown, MA 02129, USA; Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02139, USA; Howard Hughes Medical Institute, Bethesda, MD 20815, USA.
³ Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA.
⁴ Molecular Pathology Unit, Massachusetts General Hospital Research Institute, Charlestown, MA 02129, USA; Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA.
⁵ Shriners Hospitals for Children-Boston, MA 02114, USA; Center for Engineering in Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Medical School, Boston, MA 02114, USA.
⁶ Department of Pathology, Brigham and Women's Hospital, Boston, MA 02115, USA.
⁷ Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA; Howard Hughes Medical Institute, Bethesda, MD 20815, USA.
⁸ Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC 27710, USA.
⁹ Molecular Pathology Unit, Massachusetts General Hospital Research Institute, Charlestown, MA 02129, USA; Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02139, USA. Electronic address: [email protected].

Abstract

Xenograft cell transplantation into immunodeficient mice has become the gold standard for assessing pre-clinical efficacy of cancer drugs, yet direct visualization of single-cell phenotypes is difficult. Here, we report an optically-clear prkdc^-/-, il2rga^-/- zebrafish that lacks adaptive and natural killer immune cells, can engraft a wide array of human cancers at 37°C, and permits the dynamic visualization of single engrafted cells. For example, photoconversion cell-lineage tracing identified migratory and proliferative cell states in human rhabdomyosarcoma, a pediatric cancer of muscle. Additional experiments identified the preclinical efficacy of combination olaparib PARP inhibitor and temozolomide DNA-damaging agent as an effective therapy for rhabdomyosarcoma and visualized therapeutic responses using a four-color FUCCI cell-cycle fluorescent reporter. These experiments identified that combination treatment arrested rhabdomyosarcoma cells in the G2 cell cycle prior to induction of apoptosis. Finally, patient-derived xenografts could be engrafted into our model, opening new avenues for developing personalized therapeutic approaches in the future.

Keywords: SCID; breast cancer; il2rg; immune deficient; melanoma; prkdc; rhabdomyosarcoma; xenograft; zebrafish.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Genetically Modified / genetics
Animals, Genetically Modified / immunology
Animals, Genetically Modified / metabolism*
Antineoplastic Combined Chemotherapy Protocols / pharmacology*
Female
Heterografts
Humans
K562 Cells
Male
Muscle Neoplasms* / drug therapy
Muscle Neoplasms* / immunology
Muscle Neoplasms* / metabolism
Muscle Neoplasms* / pathology
Neoplasm Transplantation
Phthalazines / pharmacology
Piperazines / pharmacology
Rhabdomyosarcoma* / drug therapy
Rhabdomyosarcoma* / immunology
Rhabdomyosarcoma* / metabolism
Rhabdomyosarcoma* / pathology
Temozolomide / pharmacology
Xenograft Model Antitumor Assays
Zebrafish / genetics
Zebrafish / immunology
Zebrafish / metabolism*

Substances

Phthalazines
Piperazines
olaparib
Temozolomide

Abstract

Publication types

MeSH terms

Substances

Grants and funding