Header Forschung

Development of CRISPR/Cas9 technology for personalized, precision oncology

Summary of the Project

Despite intensive research during the last decades, treatment regimes for many cancers (including acute myeloid leukemia (AML)) have not changed significantly. Reasons for this are manifold, but a main reason is the marked genetic heterogeneity of cancers, which complicates the development of novel therapeutics. Cancer cells carry a unique, patient-specific mutation signature and it is often unclear, which mutations contribute to the disease and which ones are mere passengers without affecting malignancy. Identification of all mutations in an actual tumor of a patient by whole genome sequencing is rapidly emerging as the method of choice for precision diagnostics. However, detailed knowledge of the functional roles and relevance of most mutations arising during tumorigenesis are still lacking.

We have developed a system to functionally investigate the role and relevance of cancer mutations applying CRISPR/Cas9 technology. With this application, we propose to further develop this system to allow personalized cancer mutation profiling. We will design and produce lentiviral sgRNA libraries targeting the majority of mutations found in specific cancer cells. Functionally relevant mutations will be revealed by next generation sequencing. In a first test case we will employ an established AML cell line that will be followed by profiling samples isolated directly from AML patients. The method could become a key technology in personalized oncology with the potential to also transform individualized cancer therapy.

Scientific Goals

  • Develop a technology that can rapidly pinpoint relevant mutations in cancer patient samples.
  • Develop the CRISPR/Cas9 system as a tool that will be useful to assist physicians to identify the best treatment options for individualized treatments of their patients.
Illustration of targeting cancer mutations utilizing the CRISPR/Cas9 system.

Illustration of targeting cancer mutations utilizing the CRISPR/Cas9 system.

Copyright: Frank Buchholz

Gebler C.; Lohoff T.; Paszkowski-Rogacz M.; Mircetic J.; Chakraborty D.; Camgoz A, Hamann M.V.; Theis M.; Thiede C.; Buchholz F. Inactivation of Cancer Mutations Utilizing CRISPR/Cas9. J Natl Cancer Inst. 2016 Aug 30;109(1). pii: djw183. doi: 10.1093/jnci/djw183.

Karimova, M.; Beschorner, N.; Dammermann, W.; Chemnitz J.; Indenbirken D.; Bockmann
J.H.; Grundhoff A.; Lüth S.; Buchholz F.; Schulze zur Wiesch J.; Hauber J. CRISPR/Cas9 nickase mediated disruption of hepatitis B virus open reading frame S and X. Scientific reports. 2015, 5, 13734.

Karpinski J, Hauber I, Chemnitz J, Schäfer C, Paszkowski-Rogacz M, Chakraborty D, Beschorner N, Hofmann-Sieber H, Lange UC, Grundhoff A, Hackmann K, Schrock E, Abi-Ghanem J, Pisabarro MT, Surendranath V, Schambach A, Lindner C, van Lunzen J, Hauber J, Buchholz F. Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity. Nat Biotechnol. 2016 Apr;34(4):401-9. doi: 10.1038/nbt.3467.

Karimova, M.; Abi-Ghanem, J.; Berger, N.; Surendranath V.; Pisabarro M.T.; Buchholz F. Vika/vox, a novel efficient and specific Cre/loxP-like site-specific recombination system. Nucleic Acids Res. 2013, 41, e37.

Sarkar, I.; Hauber, I.; Hauber, J.; Buchholz F. HIV-1 proviral DNA excision using an evolved
recombinase. Science. 2007, 316, 1912-1915.

Contact

Prof. Frank Buchholz
Head of Laboratory, Medical Systems Biology
Phone: +49 351 46340288
Email: frank.buchholz(at)tu-dresden.de