Cytochrome P450 2W1 (CYP2W1) as a novel drug target in colon cancer therapy
Author: Travica, Sandra
Date: 2013-09-24
Location: Farmakologens föreläsningssal , Nanna Svartz väg 2, Karolinska Institutet, Solna.
Time: 09:00
Department: Inst för fysiologi och farmakologi / Dept of Physiology and Pharmacology
Abstract
Cytochrome P450 2W1 (CYP2W1), is a monooxygenase enzyme endogenously expressed in fetal colon and normally silenced after birth. In adult life, the expression of CYP2W1 occurs exclusively in tumor cells yielding high amounts of the enzyme in 30% of human colon cancers. Remaining the third most commonly diagnosed malignancy in the world, the colon cancer requires new chemotherapeutic strategies that would provide higher selectivity towards transformed cells. The unique confinement of CYP2W1 to tumor tissue opens the possibility for its targeting in colon cancer therapy. The studies in this thesis have focused on characterization of the CYP2W1 enzyme and the development of novel anticancer prodrugs designed for CYP2W1-specific activation.
In cancer tissues and transfected cells CYP2W1 gives multiple immunoreactive bands, suggesting that the protein might be subjected to posttranslational modifications. Our in vitro and in vivo studies reveal that CYP2W1 undergoes glycosylation at Asn177, which is enabled by the unique inverted topology of the protein in the ER membrane. Regardless of the reversed emplacement, CYP2W1 retains its catalytic function in cell systems and is capable of converting inactive substrates to potent cytotoxic species, demonstrating the potential for prodrug activation.
With the goal to develop antitumor prodrugs for specific activation by CYP2W1, we have employed a library of novel duocarmycin-based compounds and identified ICT2705 and ICT2706 as the first molecules whose cytotoxic activity is dependent on CYP2W1 enzyme. Our studies reveal the results of the first attempt of CYP2W1 targeting in cancer, both in vitro and in vivo. We show that the colon cancer cell lines expressing CYP2W1 suffer substantial DNA damage and rapid loss of viability following incubation with ICT2705 and ICT2706. Moreover, we demonstrate that the CYP2W1-positive human xenografts undergo complete growth inhibition in mice dosed with ICT2706, with no apparent deleterious effects detected in any of the treated animals. In addition, we present in vitro evidence that the potent cytotoxic effect is most likely propagated by a bystander killing mechanism.
Polymorphisms in CYP2W1 gene yield two alleles of significant frequencies, designated CYP2W1*2 and CYP2W1*6. The resulting variant enzymes carry missense mutations that might affect the therapeutic outcome of prodrug targeting as well as the course of the metastatic disease. A recent study has associated CYP2W1*2 with a decreased risk for colon cancer, suggesting the altered catalytic activity of this enzyme variant. Using the novel duocarmycin substrates, we were able to evaluate the catalytical properties of the variant enzymes for the first time. In contrast to previous findings, our studies show comparable catalytic capacity for all CYP2W1 enzymes, and no association between CYP2W1*2 genotype and colon cancer risk based on 10-fold larger patient cohorts.
In conclusion, we have found that CYP2W1 is a glycosylated enzyme with reversed ER topology, whose catalytic activity is sustained in an intact cell system and unaffected by the most frequent polymorphic changes. We have developed first prodrugs for CYP2W1 targeting in cancer and demonstrated their antitumor efficacy in a preclinical setting. The work presented herein provides the basis for a novel therapeutic approach in colon cancer chemotherapy.
In cancer tissues and transfected cells CYP2W1 gives multiple immunoreactive bands, suggesting that the protein might be subjected to posttranslational modifications. Our in vitro and in vivo studies reveal that CYP2W1 undergoes glycosylation at Asn177, which is enabled by the unique inverted topology of the protein in the ER membrane. Regardless of the reversed emplacement, CYP2W1 retains its catalytic function in cell systems and is capable of converting inactive substrates to potent cytotoxic species, demonstrating the potential for prodrug activation.
With the goal to develop antitumor prodrugs for specific activation by CYP2W1, we have employed a library of novel duocarmycin-based compounds and identified ICT2705 and ICT2706 as the first molecules whose cytotoxic activity is dependent on CYP2W1 enzyme. Our studies reveal the results of the first attempt of CYP2W1 targeting in cancer, both in vitro and in vivo. We show that the colon cancer cell lines expressing CYP2W1 suffer substantial DNA damage and rapid loss of viability following incubation with ICT2705 and ICT2706. Moreover, we demonstrate that the CYP2W1-positive human xenografts undergo complete growth inhibition in mice dosed with ICT2706, with no apparent deleterious effects detected in any of the treated animals. In addition, we present in vitro evidence that the potent cytotoxic effect is most likely propagated by a bystander killing mechanism.
Polymorphisms in CYP2W1 gene yield two alleles of significant frequencies, designated CYP2W1*2 and CYP2W1*6. The resulting variant enzymes carry missense mutations that might affect the therapeutic outcome of prodrug targeting as well as the course of the metastatic disease. A recent study has associated CYP2W1*2 with a decreased risk for colon cancer, suggesting the altered catalytic activity of this enzyme variant. Using the novel duocarmycin substrates, we were able to evaluate the catalytical properties of the variant enzymes for the first time. In contrast to previous findings, our studies show comparable catalytic capacity for all CYP2W1 enzymes, and no association between CYP2W1*2 genotype and colon cancer risk based on 10-fold larger patient cohorts.
In conclusion, we have found that CYP2W1 is a glycosylated enzyme with reversed ER topology, whose catalytic activity is sustained in an intact cell system and unaffected by the most frequent polymorphic changes. We have developed first prodrugs for CYP2W1 targeting in cancer and demonstrated their antitumor efficacy in a preclinical setting. The work presented herein provides the basis for a novel therapeutic approach in colon cancer chemotherapy.
List of papers:
I. Alvin Gomez, Jana Nekvindova, Sandra Travica, Mi‐young Lee, Inger Johansson, David Edler, Souren Mkrtchian and Magnus Ingelman‐Sundberg (2009). Colorectal cancer ‐ specific cytochrome P450 2W1: intracellular localization, glycosylation, and catalytic activity. Mol Pharmacol. 78(6):1004‐11.
Fulltext (DOI)
Pubmed
View record in Web of Science®
II. Helen M. Sheldrake, Sandra Travica, Inger Johansson, Paul M. Loadman, Mark Sutherland, Lina Elsalem, Nicola Illingworth, Alexander J. Cresswell, Tristan Reuillon, Steven D. Shnyder, Souren Mkrtchian, Mark Searcey, Magnus Ingelman‐Sundberg, Laurence H. Patterson, Klaus Pors (2013). Re‐engineering of the duocarmycin structural architecture enables bioprecursor development targeting CYP1A1 and CYP2W1 for biological activity. J Med Chem. 56(15):6273–6277.
Fulltext (DOI)
Pubmed
View record in Web of Science®
III. Sandra Travica, Klaus Pors, Paul M. Loadman, Steven D. Shnyder, Inger Johansson, Mohammed N. Alandas, Helen M. Sheldrake, Souren Mkrtchian, Laurence H. Patterson, Magnus Ingelman‐Sundberg (2013). Colon cancer‐specific cytochrome P450 2W1 converts duocarmycin analogues into potent tumor cytotoxins. Clin Cancer Res. 19(11):2952‐2961.
Fulltext (DOI)
Pubmed
View record in Web of Science®
IV. Kristina Stenstedt, Sandra Travica, Jia Guo, Isabel Barragan, Klaus Pors, Laurence Patterson, David Edler, Souren Mkrtchian, Inger Johansson, Magnus Ingelman‐Sundberg (2013). Cytochrome P450 2W1 polymorphism: functional aspects and relation to risk for colorectal cancer. [Accepted]
Fulltext (DOI)
Pubmed
View record in Web of Science®
I. Alvin Gomez, Jana Nekvindova, Sandra Travica, Mi‐young Lee, Inger Johansson, David Edler, Souren Mkrtchian and Magnus Ingelman‐Sundberg (2009). Colorectal cancer ‐ specific cytochrome P450 2W1: intracellular localization, glycosylation, and catalytic activity. Mol Pharmacol. 78(6):1004‐11.
Fulltext (DOI)
Pubmed
View record in Web of Science®
II. Helen M. Sheldrake, Sandra Travica, Inger Johansson, Paul M. Loadman, Mark Sutherland, Lina Elsalem, Nicola Illingworth, Alexander J. Cresswell, Tristan Reuillon, Steven D. Shnyder, Souren Mkrtchian, Mark Searcey, Magnus Ingelman‐Sundberg, Laurence H. Patterson, Klaus Pors (2013). Re‐engineering of the duocarmycin structural architecture enables bioprecursor development targeting CYP1A1 and CYP2W1 for biological activity. J Med Chem. 56(15):6273–6277.
Fulltext (DOI)
Pubmed
View record in Web of Science®
III. Sandra Travica, Klaus Pors, Paul M. Loadman, Steven D. Shnyder, Inger Johansson, Mohammed N. Alandas, Helen M. Sheldrake, Souren Mkrtchian, Laurence H. Patterson, Magnus Ingelman‐Sundberg (2013). Colon cancer‐specific cytochrome P450 2W1 converts duocarmycin analogues into potent tumor cytotoxins. Clin Cancer Res. 19(11):2952‐2961.
Fulltext (DOI)
Pubmed
View record in Web of Science®
IV. Kristina Stenstedt, Sandra Travica, Jia Guo, Isabel Barragan, Klaus Pors, Laurence Patterson, David Edler, Souren Mkrtchian, Inger Johansson, Magnus Ingelman‐Sundberg (2013). Cytochrome P450 2W1 polymorphism: functional aspects and relation to risk for colorectal cancer. [Accepted]
Fulltext (DOI)
Pubmed
View record in Web of Science®
Institution: Karolinska Institutet
Supervisor: Ingelman-Sundberg, Magnus
Issue date: 2013-09-02
Rights:
Publication year: 2013
ISBN: 978-91-7549-222-3
Statistics
Total Visits
Views | |
---|---|
Cytochrome ...(legacy) | 1696 |
Cytochrome ... | 409 |
Total Visits Per Month
October 2023 | November 2023 | December 2023 | January 2024 | February 2024 | March 2024 | April 2024 | |
---|---|---|---|---|---|---|---|
Cytochrome ... | 1 | 2 | 0 | 1 | 1 | 1 | 0 |
File Visits
Views | |
---|---|
Travica_thesis.pdf(legacy) | 1654 |
Travica_thesis.pdf | 348 |
Spikblad.pdf(legacy) | 296 |
Spikblad.pdf | 58 |
Travica_thesis.pdf.pdf(legacy) | 1 |
Spikblad.pdf.pdf(legacy) | 1 |
Top country views
Views | |
---|---|
United States | 590 |
Germany | 483 |
Sweden | 235 |
China | 99 |
United Kingdom | 49 |
Ukraine | 28 |
Russia | 19 |
South Korea | 16 |
Mexico | 14 |
Serbia | 14 |
Top cities views
Views | |
---|---|
Kiez | 180 |
Stockholm | 92 |
Ashburn | 62 |
Beijing | 34 |
Düsseldorf | 31 |
Nürnberg | 30 |
Romeo | 30 |
Reading | 24 |
Sunnyvale | 23 |
Seoul | 16 |