Development of novel CDK inhibitor-based combination therapy for cancer treatment
Prosjekt
- Prosjektnummer
- 2012012
- Ansvarlig person
- Jorrit Enserink
- Institusjon
- Oslo universitetssykehus HF
- Prosjektkategori
- Doktorgradsstipend
- Helsekategori
- Blood, Cancer
- Forskningsaktivitet
- 1. Underpinning, 5. Treatment Developement
Rapporter
For certain hematological cancers, such as AML and ALL, there exists a need for new forms of treatment. Furthermore, while very good treatments have been developed for certain forms of leukemia, such as CML, development of resistance to these treatments continues to be a problem. The goal of this project was to develop novel forms of therapy for leukaemia.
We screened for compounds that selectively kill leukemia cells. We identified several such compounds. One of these was a novel compound that was not previously known to selectively target oncogene-expressing cancer cells. We have studied the mechanism of action of this compound and discovered that it has several effects on cancer cells, but the most notable effect is that it induces rapid loss of the transcription factor c-Myc. Most -if not all- cancer cells require c-Myc for their proliferation and survival, making c-Myc an attractive target for therapy. However, since it lacks catalytic activity, it is difficult to design molecules that directly target c-Myc. We discovered that our compound blocks the synthesis of c-Myc at the level of mRNA synthesis/stability.
Importantly, our compound is particularly toxic for leukemic stem cells, which constitute a small subpopulation of cells that can cause disease relapse in patients who fail standard of care treatment. Finally, the compound is not only active against leukemias, but other forms of cancer appear to be sensitive to the compound as well. We are currently in the process of preparing two manuscripts for publication, and a PhD thesis has been submitted to the Medical Faculty of the University of Oslo, which we expect will result in a PhD degree by the middle of this year.
This project was an early-stage basic research project aimed at identifying novel compounds that may be used for cancer treatment, and will therefore not have immediate impact on health services. However, we expect that the research methods that we developed in this project, as well as the bioinformatics tools, may assist follow-up studies that in the longer term have more direct effects on health services.
Nei
For certain hematological cancers, such as AML and ALL, there exists a need for new forms of treatment. Furthermore, while good treatments have been developed for CML, development of resistance to these treatments continues to be a problem. The goal of this project was to develop of novel forms of therapy.KML og en viss form av ALL ("Ph+ ALL") er forårsaket av onkogen BCR-Abl. BCR-Abl kan være effektivt hemmet av små molekyl hemmere som imatinib (Glivec) og flere neste generasjons BCR-ABL-hemmere. KML-pasienter reagerer svært godt til disse stoffene og har en forventet levealder som kan sammenlignes med friske mennesker. Ph+ ALL pasienter derimot reagerer ikke så godt til disse stoffene, og nye terapistrategier er nødvendig for behandling av disse pasientene.
Vi har utviklet en ny metode for å raskt screene for medikamenter som spesifikt dreper BCR-ABL-uttrykkende celler men ikke friske celler. Vi har undersøkt nesten 20.000 forbindelser og har identifisert flere stoffer som effektivt dreper Ph+ ALL celler. Vi undersøker nå deres virkningsmekanisme ved hjelp av en kombinasjon av moderne teknologier. For eksempel, vi prøver å forstå hvordan disse forbindelsene påvirker aktiviteten av intracellulære signalveier, og vi er i fart ved å kartlegge spesifikke fosforyleringer med bruk av bruker fosfor-massespektrometri. Ved hjelp av denne fremgangsmåten har vi funnet at i det minste en av disse forbindelser kan spesifikt hemme transkripsjonsfaktoren Myc. Myc spiller en avgjørende rolle i å drive tumorvekst i mange former for kreft. Vi ønsker nå å forstå den molekylære mekanismen som ligger bak inaktivering av Myc, og våre foreløpige data tyder på at denne forbindelsen destabiliserer Myc mRNA. Vi har påvist at dette molekylet er aktiv mot celler fra kreftpasienter, og vi er nå gang med dyreforsøk for å teste om dette molekylet er også aktiv mot kreft i et mus modell av kreft. I tillegg så har vi sendt inn en patentsøknad.
KML og en viss form av ALL ("Ph+ ALL") som er forårsaket av onkogen BCR-Abl. KML og Ph+ ALL behandles med små molekyl hemmere som imatinib. Overlevelsen til Ph+ ALL pasienter er fremdeles ikke optimalt, og nye terapistrategier er nødvendig for behandling av disse pasientene. Vi har identifisert potensielle nye behandlingsstrategier.KML og en viss form av ALL ("Ph+ ALL") er forårsaket av onkogen BCR-Abl. BCR-Abl kan være effektivt hemmet av små molekyl hemmere som imatinib (Glivec/Gleevec) og flere neste generasjons BCR-ABL-hemmere. KML-pasienter reagerer svært godt til disse stoffene og har en forventet levealder som kan sammenlignes med friske mennesker. Ph+ ALL pasienter derimot reagerer ikke så godt til disse stoffene, og nye terapistrategier er nødvendig for behandling av disse pasientene. Vi har utviklet en ny metode for å raskt screene for medikamenter som spesifikt dreper BCR-ABL-uttrykkende celler men ikke friske celler. Vi har undersøkt nesten 20.000 forbindelser og har identifisert flere stoffer som effektivt dreper Ph+ ALL celler. Akkurat nå undersøker vi virkningsmekanisme til disse stoffene ved hjelp av en kombinasjon av moderne teknologier. For eksempel, vi ønsker å forstå hvordan disse forbindelsene påvirker aktiviteten av intracellulære signalveier, og vi har kartlagt spesifikke fosforyleringer med bruk av fosfomassespektrometri. Ved hjelp av denne fremgangsmåten har vi funnet at i det minste en av disse forbindelser kan spesifikt hemme transkripsjonsfaktoren Myc. Myc spiller en avgjørende rolle i å drive tumorvekst i mange former for kreft. Vi prøver nå å forstå den molekylære mekanismen som bak inaktivering av Myc, og våre foreløpige data tyder på at denne forbindelsen aktiverer en fosfatase som dephosphorylerer og dermed inaktiverer Myc. I tillegg har vi funnet ut at en av de andre stoffene induserer nedbryting av flere onkogener, blant annet BCR-Abl. Vi har som mål å videreutvikle disse stoffene og utprøve deres aktivitet mot celler fra kreftpasienter.
- KML og en viss form av ALL ("Ph+ ALL") er forårsaket av onkogen BCR-Abl
- KML behandles med små molekyl hemmere som imatinib
- Ph+ ALL pasienter derimot reagerer ikke så godt til disse stoffene, og nye terapistrategier er nødvendig for behandling av disse pasientene
- Vi har identifisert en små molekyl hemmer av c-MycKML og en viss form av ALL ("Ph+ ALL") er forårsaket av onkogen BCR-Abl. BCR-Abl kan være effektivt hemmet av små molekyl hemmere som imatinib (Glivec) og flere neste generasjons BCR-ABL-hemmere. KML-pasienter reagerer svært godt til disse stoffene og har en forventet levealder som kan sammenlignes med friske mennesker. Ph+ ALL pasienter derimot reagerer ikke så godt til disse stoffene, og nye terapistrategier er nødvendig for behandling av disse pasientene.
Vi har utviklet en ny metode for å raskt screene for medikamenter som spesifikt dreper BCR-ABL-uttrykkende celler men ikke friske celler. Vi har undersøkt nesten 20.000 forbindelser og har identifisert flere stoffer som effektivt dreper Ph+ ALL celler. Vi undersøker nå deres virkningsmekanisme ved hjelp av en kombinasjon av moderne teknologier. For eksempel, vi prøver å forstå hvordan disse forbindelsene påvirker aktiviteten av intracellulære signalveier, og vi er i fart ved å kartlegge spesifikke fosforyleringer med bruk av bruker fosfor-massespektrometri. Ved hjelp av denne fremgangsmåten har vi funnet at i det minste en av disse forbindelser kan spesifikt hemme transkripsjonsfaktoren Myc. Myc spiller en avgjørende rolle i å drive tumorvekst i mange former for kreft. Vi prøver nå å forstå den molekylære mekanismen som bak inaktivering av Myc, og våre foreløpige data tyder på at denne forbindelsen aktiverer en fosfatase som dephosphorylerer og dermed inaktiverer Myc. Vi har også som mål å videreutvikle denne forbindelsen og utprøve sin aktivitet mot celler fra kreftpasienter.
Hvert år omtrent 1000 nye tilfeller av leukemi er diagnostisert i Norge . Flere av disse kreftformer, spesielt KML og en viss form for ALL, er forårsaket av onkogen BCR-Abl. ALL pasienter responderer relativt dårlig til dagens standardbehandling, og det trengs nye former for behandling av disse pasienter.Norsk:
KML og en viss form av ALL ("Ph+ ALL") er forårsaket av onkogen BCR-Abl. BCR-Abl kan være effektivt hemmet av små molekyl hemmere som imatinib (Glivec) og flere neste generasjons BCR-ABL-hemmere. KML-pasienter reagerer svært godt til disse stoffene og har en forventet levealder som kan sammenlignes med friske mennesker. Ph+ ALL pasienter derimot reagerer ikke så godt til disse stoffene, og nye terapistrategier er nødvendig for behandling av disse pasientene.
Vi har utviklet en ny metode for å raskt screene for medikamenter som spesifikt dreper BCR-ABL-uttrykkende celler men ikke friske celler. Vi har undersøkt nesten 20.000 forbindelser og har identifisert flere stoffer som effektivt dreper Ph+ ALL celler. Vi undersøker nå deres virkningsmekanisme ved hjelp av en kombinasjon av moderne teknologier. For eksempel, vi prøver å forstå hvordan disse forbindelsene påvirker aktiviteten av intracellulære signalveier, og vi er i fart ved å kartlegge spesifikke fosforyleringer med bruk av bruker fosfor-massespektrometri. Ved hjelp av denne fremgangsmåten har vi funnet at i det minste en av disse forbindelser kan spesifikt hemme transkripsjonsfaktoren Myc. Myc spiller en avgjørende rolle i å drive tumorvekst i mange former for kreft. Vi prøver nå å forstå den molekylære mekanismen som bak inaktivering av Myc, og våre foreløpige data tyder på at denne forbindelsen aktiverer en fosfatase som dephosphorylerer og dermed inaktiverer Myc. Vi har også som mål å videreutvikle denne forbindelsen og utprøve sin aktivitet mot celler fra kreftpasienter.
English:
CML and a certain form of ALL ("Ph+ ALL") are caused by the oncogene BCR-Abl. BCR-Abl can be effectively targeted by small molecule inhibitors like imatinib (gleevec) and several next-generation BCR-Abl inhibitors. CML patients respond very well to these drugs and have a life expectancy comparable to healthy people. However, Ph+ ALL patients do not respond that well to these drugs, and new forms of treatment are required for treatment of these patients.
We have developed a novel method to rapidly screen for drugs that specifically kill BCR-Abl-expressing cells, but not healthy cells. We screened nearly 20,000 compounds and identified several drugs that effectively kill Ph+ ALL cells. We are now investigating their mechanism of action using a combination of modern technologies. For instance, we are trying to understand how these compounds affect the activity of intracellular signaling pathways by mapping phosphorylation sites using phospho-mass spectrometry. Using this method we found that at least one of these compounds may specifically inhibit the transcription factor Myc. Myc plays a crucial role in driving tumor growth in many forms of cancer. We are now trying to elucidate the molecular mechanism by which this compound inactivates Myc, and our preliminary data indicate that this compound activates a phosphatase which in turn dephosphorylates and thereby inactivates Myc. We also aim to further develop this compound and test its activity against cells derived from cancer patients.
The cyclin dependent kinase CDK1 is the master regulator of the cell cycle. It is aberrantly activated in all cancers, and therefore it is believed that drugs that target CDK1, as well as related proteins like CDK2, CDK4 and CDK6, may be effective against various forms of cancer.The cyclin dependent kinase CDK1 is the master regulator of the cell cycle. It regulates the activity of a number of proteins involved in cell cycle-dependent processes, including DNA replication, DNA repair and mitosis. CDK1, and sometimes related CDKs such as CDK2, CDk4 and CDK6, are aberrantly activated in all cancers, and therefore it is believed that drugs that target CDK1, CDK2, CDK4 or CDK6, may be effective against various forms of cancer. However, results from clinical trials with CDK inhibitors, such as flavopyridol, have generally been disappointing. This is mainly due to general toxicity and lack of efficacy. More recent clinical trials with the CDK4/CDK6 inhibitors have shown promising results for treatment of breast cancer.
It is now well understood that combination treatment often has higher efficacy than treatment of patients with individual drugs. For instance, younger AML patients, such as those under 60, are typically treated with two chemo drugs, cytarabine and an anthracycline drug such as daunorubicin (daunomycin) or idarubicin. Sometimes a third drug, cladribine (Leustatin, 2-CdA), is given as well. We reasoned that perhaps CDK inhibitors by themselves are not very potent anti-cancer drugs, but when combined with other drugs they may show potent activity against cancer cells.
We devised a chemical-genetic cell screen to identify drugs that synergize with CDK inhibitors. This screen identified a number of candidate compounds that cooperate with CDK inhibitors. We are currently studying the mechanism by which these drug combinations synergize.
De cyclin avhangige kinasene (CDKs) er sentrale regulatorer av celle syklusen
Disse enzymene passer på at visse kontroll punkter i cellesyklusen overholdes. Desverre har flertallet av kreft cellene mistet kontrollen over sine CDKs og dette gir disse cellene et tet vekst fortinn over de andre cellene i det aktuelle vevet.Cell proliferation is regulated by cyclin dependent kinases (CDKs), which are serine/threonine protein kinases whose activity depends on binding and activation by cyclin partners. These heterodimeric complexes phosphorylate various substrates to control cell cycle progression in response to different stimuli. Several CDKs are involved in cell cycle regulation in mammalian cells, and while a single CDK (CDK1) is sufficient for basic cell cycle control, CDK2, CDK4 and CDK6 are required for cell cycle regulation of specialized cells.
Importantly, the activity of CDKs is aberrantly increased in all forms of cancer, due to the frequent overexpression of positive regulators (e.g. cyclin D) and the frequent inactivation of CDK inhibitors (p21, p27, etc). Abnormalities in CDK activity have also been found in viral infections, proliferative renal diseases and neurodegenerative disorders such as Alzheimer's, Parkinson's and Nieman-Pick's disease, ischemia and traumatic brain injury. Not surprisingly, this has sparked an intensive search for small-molecule CDK inhibitors for therapeutic applications. However, no CDK inhibitor has currently been approved for commercial use. Early clinical compounds, such as the pan-CDK inhibitors flavopiridol and CY-202 have undergone numerous phase II and phase III clinical trials. The results of these trials have been disappointing, although flavopiridol recently demonstrated activity in individuals with refractory chronic lymphocytic leukemia.
Several second-generation CDK inhibitors that are considerably more specific are currently in clinical trial with promising preliminary results. Furthermore, combination therapy, in which two drugs synergize to block cell proliferation/survival, has the additional benefit that lower doses of drugs can be used, circumventing the problem of general toxicity.
To identify drugs that might be used in combination therapy, we recently performed a high-throughput drug screen for compounds that synergize with pharmacological CDK inhibitors. This screen identified several drugs that, when combined with specific CDK inhibitors, result in decreased cell survival. One of these drugs targets p38MAPK. p38MAPK is a well-know stress-induced kinases with multiple functions in the cell. In some cell types activation of p38 results in increased cell death whereas in other cell types it promotes apoptosis. We found small-molecule p38MAPK inhibitors synergize with CDK inhibitors to induce apoptosis in an oncogenic B cell line. We are currently dissecting the molecular mechanism that causes the high rates of apoptosis.
Many forms of cancer remain poorly treatable, and there exists a need for a novel form of chemotherapy that inhibits growth of tumor cells that do not respond to first line chemotherapies, that is less toxic and that does not carry the risk of inducing secondary tumors.Cell proliferation is regulated by cyclin dependent kinases (CDKs), which are serine/threonine protein kinases whose activity depends on binding and activation by cyclin partners. Importantly, the activity of CDKs is aberrantly increased in all forms of cancer, due to the frequent overexpression of positive regulators (e.g. cyclin D) and the frequent inactivation of CDK inhibitors.
Not surprisingly, this has sparked an intensive search for small-molecule CDK inhibitors for therapeutic applications. However, no CDK inhibitor has currently been approved for commercial use due to low specificity of some of these drugs, leading to general toxicity.
Cell proliferation is regulated by cyclin dependent kinases (CDKs), which are serine/threonine protein kinases whose activity depends on binding and activation by cyclin partners. These heterodimeric complexes phosphorylate various substrates to control transcription and cell cycle progression in response to different stimuli. In mammalian cells several CDKs are involved in cell cycle regulation, and while a single CDK (CDK1) is sufficient for basic cell cycle control, CDK2, CDK4 and CDK6 are required for cell cycle regulation of specialized cells.
Importantly, the activity of CDKs is aberrantly increased in all forms of cancer, due to the frequent overexpression of positive regulators (e.g. cyclin D) and the frequent inactivation of CDK inhibitors (p21, p27, etc). Abnormalities in CDK activity have also been found in viral infections, proliferative renal diseases and neurodegenerative disorders such as Alzheimer's, Parkinson's and Nieman-Pick's disease, ischemia and traumatic brain injury. Not surprisingly, this has sparked an intensive search for small-molecule CDK inhibitors for therapeutic applications. However, no CDK inhibitor has currently been approved for commercial use. Early clinical compounds, such as the pan-CDK inhibitors flavopiridol and CY-202 have undergone numerous phase II and phase III clinical trials. The results of these trials have been disappointing, although flavopiridol recently demonstrated activity in individuals with refractory chronic lymphocytic leukemia.
The low efficacy of the initial CDK inhibitors may be the result of several problems, like low specificity of some of these drugs, leading to general toxicity. The genetic background of the tumor cells also determines whether they may be sensitive to CDK inhibitors. For example, tumors that are driven by mutations downstream of CDKs, like e.g. mutations in the Retinoblastoma gene RB, are not likely to be susceptible to CDK inhibitors because the mutation in RB may (at least partially) bypass the dependence on CDK activity. However, the majority of tumors are driven by mutations that increase CDK activity, and those tumors are likely to be sensitive to CDK inhibitors. Several second-generation CDK inhibitors that are considerably more specific are currently in clinical trial, and combination therapy, in which CDK inhibitors are combined with other pharmaceutical compounds, may result in much better clinical response.
We recently performed a high-throughput chemical-genetic screen to search for compounds that synergize with CDK inhibitors in reducing cell proliferation/survival. This screen identified five lead compounds that by themselves did not affect cell proliferation, but when combined with low (sub-IC50) doses of a CDK inhibitor they potently inhibited cell survival/proliferation. The goal of this project proposal is to further characterize the effect of combining CDK inhibitors and these lead compounds on cell proliferation and survival. The ultimate aim is to develop the results of these pre-clinical studies into a combination chemotherapy to treat cancer patients.
Vitenskapelige artikler
Pietarinen, Eide, Ayuda-Durán, Potdar, Kuusanmäki, Andersson, Mpindi, Pemovska, Kontro, Heckman, Kallioniemi, Wennerberg, Hjorth-Hansen, Druker, Enserink, Tyner, Mustjoki, and Porkka
Differentiation status of primary chronic myeloid leukemia cells affects sensitivity to BCR-ABL1 inhibitors
Oncotarget, 2017 in press
Deltagere
- Marketa Chlubnova Prosjektdeltaker
- Richard Crispin Prosjektdeltaker
- Emmet Mc Cormack Prosjektdeltaker
- Sebastien Waelchli Prosjektdeltaker
- Dagim Shiferaw Tadele Postdoktorstipendiat (finansiert av denne bevilgning)
- Jorrit Enserink Prosjektleder
- Beibei Zhang Postdoktorstipendiat (annen finansiering)
- Joseph Robertson Postdoktorstipendiat (annen finansiering)
- Laure Piechaczyk Doktorgradsstipendiat (annen finansiering)
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