Abstract
Thapsigargin and nortrilobolide are sesquiterpene lactones found in the Mediterranean plant Thapsia
garganica L. Thapsigargin is a potent inhibitor of the sarco/endoplasmic reticulum calcium ATPase
(SERCA) pump, inducing apoptosis in mammalian cells. Due to its ability to induce apoptosis, thapsigargin
has become the active part of a pro-drug (Mipsagargin) for the treatment of different cancer types producing
promising results in patients with hepatocellular carcinoma, glioblastoma, prostate cancer and renal cell
carcinoma. The thapsigargin-based prodrug has been patented by the American company Inspyr
Therapeutics under the name of Mipsagargin. The annual demand for thapsigargin is expected to be 1 ton per
year; therefore, there is an increasing need to discover the thapsigargin biosynthesis; and to develop novel
production platforms that allow for large scale production.
In this PhD project, I present a review article about the evolution of the bioactive compound thapsigargin
produced by Thapsia garganica until the development of the prodrug Mipsagargin (chapter I). Next, in
chapter II I have established an alternative production platform for thapsigargin production based on in vitro
tissue cultures. In this chapter has been described, from the regeneration of in vitro plants of T. garganica, to
the establishment and enhancement of thapsigargins production in temporary immersion bioreactors. A part
from an alternative production platform for thapsigargins, in vitro plant tissue cultures techniques have been
described for this medicinal species, which can be used for plant genetic conservation and biodiversity.
In the chapters II, III and IV, I present studies on the expression levels of the two biosynthetic genes
described in the thapsigargin biosynthesis, TgTPS2 and TgCYP76AE2. These studies were performed with
T. garganica in vitro plants under stress treatments, with plants growing in the greenhouse and with wild
plants. We wanted to understand the complex mechanisms determining the production of thapsigargins in
order to transpose this knowledge for futures approaches.
Finally, I was encouraged to genetically transform T. garganica (chapter V). I tried to design an
Agrobacterium tumefaciens-mediated transformation protocol. Nevertheless, the performed experiments did
not work. It was evident that a bigger investment in time is necessary to achieve Thapsia transformation, but
I am confident that it is possible.
garganica L. Thapsigargin is a potent inhibitor of the sarco/endoplasmic reticulum calcium ATPase
(SERCA) pump, inducing apoptosis in mammalian cells. Due to its ability to induce apoptosis, thapsigargin
has become the active part of a pro-drug (Mipsagargin) for the treatment of different cancer types producing
promising results in patients with hepatocellular carcinoma, glioblastoma, prostate cancer and renal cell
carcinoma. The thapsigargin-based prodrug has been patented by the American company Inspyr
Therapeutics under the name of Mipsagargin. The annual demand for thapsigargin is expected to be 1 ton per
year; therefore, there is an increasing need to discover the thapsigargin biosynthesis; and to develop novel
production platforms that allow for large scale production.
In this PhD project, I present a review article about the evolution of the bioactive compound thapsigargin
produced by Thapsia garganica until the development of the prodrug Mipsagargin (chapter I). Next, in
chapter II I have established an alternative production platform for thapsigargin production based on in vitro
tissue cultures. In this chapter has been described, from the regeneration of in vitro plants of T. garganica, to
the establishment and enhancement of thapsigargins production in temporary immersion bioreactors. A part
from an alternative production platform for thapsigargins, in vitro plant tissue cultures techniques have been
described for this medicinal species, which can be used for plant genetic conservation and biodiversity.
In the chapters II, III and IV, I present studies on the expression levels of the two biosynthetic genes
described in the thapsigargin biosynthesis, TgTPS2 and TgCYP76AE2. These studies were performed with
T. garganica in vitro plants under stress treatments, with plants growing in the greenhouse and with wild
plants. We wanted to understand the complex mechanisms determining the production of thapsigargins in
order to transpose this knowledge for futures approaches.
Finally, I was encouraged to genetically transform T. garganica (chapter V). I tried to design an
Agrobacterium tumefaciens-mediated transformation protocol. Nevertheless, the performed experiments did
not work. It was evident that a bigger investment in time is necessary to achieve Thapsia transformation, but
I am confident that it is possible.
| Originalsprog | Engelsk |
|---|
| Forlag | Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen |
|---|---|
| Status | Udgivet - 2017 |