Molecular Oncology

The laboratory studies the cellular mechanisms and genetic alterations involved in human cancer to better understand cancer, establish better classifications, identify stem cells and mutated genes, and identify diagnostic and prognostic markers that can be transferred into routine use. We study various types of human tumors.

In breast cancers, the teams of François Bertucci, Max Chaffanet/Anne Letessier, Emmanuelle Charafe-Jauffret/Christophe Ginestier, and Marc Lopez work at identifying cancer stem cells, genomic alterations specific of molecular subtypes and histoclinical forms, prognostic and predictive markers, and to characterize the role and function of mutated genes. In vivo models such as transgenic and xenografted mice allow pre-clinic studies.

The team of Palma Rocchi develops new therapeutic drugs against prostate cancer.

Identification of mutated genes in chronic and acute myeloid hematopoietic diseases is done by the team of Marie-Joelle Mozziconacci, Anne Murati and Véronique Gelsi-Boyer.

The team of Geraldine Guasch studies the molecular mechanism involved in tumor formation in epithelial transition zoneswhere one type of epithelium changes to another such as the cervix, anorectal region, the esophagus-stomach and the limbus of the eye. They have identified the presence of a stem cell niche in those regions and have established a mouse model that recapitulates tumor formation in those regions.

Finally, Emilie Mamessier and Claire Acquaviva's team explores circulating tumor cell (CTCs) heterogeneity to identify cells at high risk of seeding metastases.

Recent discoveries:

  • A new oncogene in breast cancer : The ZNF703 oncogene at 8p12 is amplified in luminal B breast cancers. ZNF703 plays a role in stem cell biology.
  • One of the most frequently mutated gene in malignant myeloid blood diseases : ASXL1 is frequently mutated in myeloid diseases and encodes a regulator of polycomb genes.
  • Mutations of SWI/SNF component ARID1A in pancreatic and breast cancers.

Breast cancer

The team's effort is focused on understanding the various steps of breast carcinogenesis development. We study the hierarchical relationships between cells in normal and malignant breast epithelium in order to gain insight into the processes that underlie tumor initiation and development, and metastasis formation.

The primary aim is to characterize and to understand the regulation of mammary epithelial stem cells since these are likely to be the targets of cancer-initiating events, and may be the underlying tumorigenic cells in breast cancers.

The secondary aim is to characterize the breast cancer stem cell population of various tumor subtypes to identify new markers and therapeutic targets and use the cancer stem cell concept at the clinical level to manage the disease.

 

Getting to the root of breast cancer

The CSC model holds that tumors are organized in a cellular hierarchy in which "cancer stem cells" are the only cells with unlimited proliferation potential and with the capability of driving tumor growth and expansion. According to this model, cancers originate from the malignant transformation of an adult stem cell or progenitor through the dysregulation of the normally tightly regulated self-renewal program. This leads to clonal stem/progenitor cell expansion generating cells that then undergo further genetic or epigenetic alterations to become fully transformed. As a consequence of this, tumors contain a cellular component of "cancer stem cells" (CSC) which retain key stem cell properties that initiate and drive carcinogenesis. Understanding the molecular basis for dysregulated self-renewal is crucial for identification of targets for effective therapeutic intervention. Moreover, it will help us understand how the different molecular subtypes originate from CSC.

To study the different mechanisms that initiate oncogenesis, we perform in vitro and in vivo experiments in immunodeficient mice utilizing human mammary epithelial cells from patients that undergo reduction mammoplasties. These cells can be modified by lentiviral infection for the overexpression or downregulation of candidate genes.

 

The cancer stem cell : the breast cancer driver

The cancer stem cell hypothesis modifies our conceptual approach of oncogenesis and described the CSC as "the cell to be killed" to eradicate breast cancer. To better characterize this cell population, we develop several strategies based on genomic, transcriptomic, and proteomic profiling. One current interest is to study the ALDEFLUOR-positive population from different breast cancer subtypes to identify the different molecular mechanisms that sustain CSC biology. To validate functionally the different candidate genes/pathways identified, we develop a human breast tumor xenobank by transplantation of human primary tumor in immunodeficient mice. Moreover new therapeutic approaches targeting specifically the CSC population are evaluated utilizing these xenografts.

 

Bibliography

  1. Ginestier C, Liu S, Diebel ME, Korkaya H, Luo M, Brown M, et al.CXCR1 blockade selectively targets human breast cancer stem cells in vitro and in xenografts. J Clin Invest. 2010 Feb 120:2.
  2. Charafe-Jauffret E, Ginestier C, Iovino F, Tarpin C, Diebel M, Esterni B et al. Aldehyde Dehydrogenase 1-Positive Cancer Stem Cells Mediate Metastasis and Poor Clinical Outcome in Inflammatory Breast Cancer. Clin Cancer Res. 2009 Dec 22.
  3. Ginestier C, Wicinski J, Cervera N, Monville F, Finetti P, Bertucci F, et al. Retinoid signaling regulates breast cancer stem cell differentiation. Cell Cycle. 2009 Oct 15 ;8(20):3297-302.
  4. Ginestier C, Liu S, Wicha MS. Getting to the root of BRCA1-deficient breast cancer. Cell Stem Cell. 2009 Sep 4 ;5(3):229-30.
  5. Charafe-Jauffret E, Ginestier C, Birnbaum D. Breast cancer stem cells : tools and models to rely on. BMC Cancer. 2009 Jun 25 ;9:202. Review.
  6. Korkaya H, Paulson A, Charafe-Jauffret E, Ginestier C, Brown M, Dutcher J, et al. Regulation of mammary stem/progenitor cells by PTEN/Akt/beta-catenin signaling. PLoS Biol. 2009 Jun 2 ;7(6):e1000121.
  7. Charafe-Jauffret E, Ginestier C, Iovino F, Wicinski J, Cervera N, Finetti P, et al. Breast cancer cell lines contain functional cancer stem cells with metastatic capacity and a distinct molecular signature. Cancer Res. 2009 Feb 15 ;69(4):1302-13.
  8. Charafe-Jauffret E, Monville F, Ginestier C, Dontu G, Birnbaum D, Wicha MS. Cancer stem cells in breast : current opinion and future challenges. Pathobiology. 2008 ;75(2):75-84.
  9. Ginestier C, Hur MH, Charafe-Jauffret E, Monville F, Dutcher J, Brown M, et al. ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell. 2007 Nov ;1(5):555-67.
  10. Liu S, Ginestier C, Charafe-Jauffret E, Foco H, Kleer CG, Merajver SD, et al. BRCA1 regulates human mammary stem/progenitor cell fate. Proc Natl Acad Sci U S A. 2008 Feb 5 ;105(5):1680-5.
  11. Ginestier C, Korkaya H, Dontu G, Birnbaum D, Wicha MS, Charafe-Jauffret E. [The cancer stem cell : the breast cancer driver]. Med Sci (Paris). 2007 Dec ;23(12):1133-9. Review. French.

Transition Zones - Cancer and Stem Cells

Squamous cell carcinoma arise in stratified squamous epithelia of the body, and are one of the most common malignancies in humans with some of the poorest prognoses. They are frequently found at epithelial transition zones, where one type of epithelium changes to another such as the cervix, anorectal region, the esophagus-stomach and the limbus of the eye.
The underlying mechanisms for this increased tumor susceptibility are unknown. Our current model focuses on anorectal transitional epithelium because we have created a tumor model in mice that recapitulates human anal transition zone tumors. Our research has identified an unsuspected link between the normal stem cells of the skin and an abnormally high incidence of tumors that arise in the transitional epithelium, in that both are regulated by the TGFβ signaling pathway. Our laboratory uses the transitional epithelium as a unique model to investigate the molecular basis for how normal epithelial stem cells become cancer stem cells. Our goal in working in a medical center is to establish a research program that uses basic discoveries of stem cell biology to impact several clinical programs here including oncology.


We use a variety of molecular, biochemical and tissue culture techniques, in vivo imaging and fluorescence-activated cell sorting, as well as mouse transgenic and gene targeting (knockout) technology to address these various questions.

Current projects

1. Regulation of cancer stem cells from transition zone by their niche.

- Define the pre-tumoral niche of transition zones susceptible to develop metastatic carcinoma.
- In vitro and in vivo study of the molecular mechanisms involved in the interaction between fibroblasts and cancer stem cells at the transition zone.
- Define the properties of transition zone fibroblasts in human cancer.

2. Characterization of the molecular mechanisms involved in the metastatic properties of cancer stem cells of the transition zone.



3. Targeting of a potential mechanism activated in cancer stem cells from transition zone carcinoma.

 

Team Members

Géraldine Guasch, Ph.D CR1 Inserm

Véronique Chevrier, Ph.D IR1 Inserm

Zohra Berrehail, M2 Pathologie humaine (Arrivée Janv 2016)

Guasch Géraldine, Ph.D, CR1

 

Education and Work History


2015-Present: Tenured CR1 Inserm position in French-wide competition, Department of Molecular Oncology, CRCM, Marseille, France

2008-06/15: Assistant Professor of Pediatrics, Division of Developmental Biology, Children’s Hospital Research Foundation, Cincinnati, OH, USA

2005-2008: Postdoctoral Associate, Mammalian Cell Biology and Development
Laboratory, Rockefeller University New York, NY, USA

2002-2005: Postdoctoral Fellow, Mammalian Cell Biology and Development
Laboratory, Rockefeller University New York, NY, USA under the supervision of Dr. 
Elaine Fuchs

1998-2002: Ph.D Fellow, Molecular Oncology Laboratory, Marseille- France

1997: Undergraduate Fellow, Molecular Genetic Institute, Montpellier- France

Awards and Honors

2015-2017: Foundation ARC for Research Against Cancer Grant award
(principal Investigator)

2015-2020: NIH/NEI R01 Grant Award (co-investigator)

2014-2016: NIH/NIAMS R21 Grant Award (principal Investigator)

2011-2013: V Scholar Award (principal Investigator)

2011-2013: Sidney Kimmel Scholar Award (principal Investigator)

2011-2013: Basil’O Connor Scholar Award (principal Investigator)

2010-2012: Concern Foundation Grant Award (principal Investigator)

2009-2011: Industry Grant Award (L’Oréal) - patent PCT/US2013/063367
“Sebocyte Cell Culturing and Methods of Use” (principal inventor)

2009-2011: Trustee Grant Award, Cincinnati Children’s Hospital Medical Center

2009: SID/Eugene M. Farber Travel Award for Young Investigators, for outstanding
abstract submitted to the Symposium on ‘’Genetics-Epigenetics of Skin Diseases’’

2002: Award for the outstanding thesis of the year

2002: Foundation for Medical Research Award

2002-2005: Human Frontiers (HFSP) Postdoctoral Fellowship

2001-2002: National league against Cancer Award

1998-2001: French Ministry of Research Fellowship

 

 

Véronique Chevrier, Ph.D, IR1

 

Véronique has obtained her Ph.D in Biology in 1992 in Grenoble, France.

She is an expert in cell biology and has studied and published numerous papers on centrosome

 

 

Most relevant publications to the current projects

1. McCauley H, Guasch G. Serial orthotopic transplantation of epithelial tumors in single-cell suspension. Methods Mol Biol. 2013. 1035:231-45.


2. McNairn A., Guasch G. Epithelial Transition Zones: merging microenvironments, niches, and cellular transformation. European Journal of Dermatology. 2011, 21(Suppl.2):21-28. Review.


3. Runck L., Kramer M., Ciraolo G., Lewis A., Guasch G. Identification of epithelial label-retaining cells at the transition between the anal canal and the rectum in mice. Cell Cycle. 2010, 15: 3039-3045. Cover Article.


4. Guasch G., Schober M., Pasolli A., Conn E., Polak L., Fuchs E. Loss of TGF? signalling destabilizes homeostasis and promotes squamous cell carcinomas in stratified epithelia. Cancer Cell. 2007, 12: 313-327. Featured Article


    Highlighted by: Wakefield LM et al., Cancer Cell. 2007, 12: 313-327.


5. Tumbar T., Guasch G., Greco V., Blanpain C., Lowry W., Rendl M., Fuchs E. Defining the epithelial stem cell niche in skin. Science. 2004, 303: 359-363.


6. Fuchs E., Tumbar T., Guasch G. Socializing with the neighbors: stem cells and their niche.Cell. 2004, 116: 769-778. Review.

Additional recent publications of importance to the field (in chronological order)


7. McCauley, H.A. and Guasch G. Three Cheers For The Goblet Cell: Maintaining Homeostasis In Mucosal Epithelia. Trends In Molecular Medicine, 2015 21(8):492-503. Cover


8. McCauley H.A, Liu C-Y, Attia A, Wikenheiser-Brokamp KA, Zhang Y, Whitsett JA, and Guasch G. TGFβ signaling inhibits goblet cell differentiation via SPDEF in the conjunctival epithelium. Development. 2014 141:4628-4639.


9. Gupta A., Bischoff A., Pena A., Runck L.A., Guasch G. The Great divide: septation and malformation of the cloaca, and its implications for surgeons. Pediat Surg Int. 2014, 30(11):1089-95.


10. Runck LA, Method A, Bischoff A, Levitt M, Peña A, Collins M, Gupta A, Shanmukhappa S, Wells J.M, Guasch G. Defining the molecular pathologies in cloaca malformation: similarities between mouse and human. Dis Model Mech. 2014, 7(4):483-93.

Join the Lab

Applications for PhD scholarships and Master students are considered. Post-doctoral applicants with the possibility of applying for fellowships (HFSP, EU Marie Curie and EMBO) are welcome to apply. Please send a CV to geraldine.guasch-grangeon(at)inserm(dot)fr

 

 

Prostate cancer

Prostate cancer (PC) represents one of the most common cancers in industrialized countries.

Patients with localized disease may be treated with surgery or radiation, while androgen ablation is used as first- line therapy in patients with metastatic disease. While most patients initially respond well to this hormonal therapy, they most ultimately become unresponsive and recur within 2 years as castration- resistant prostate cancer (CRPC).

Recently, docetaxel-based regimens have demonstrated improved survival in men with CRPC in two different, large, phase III studies. However, the median overall survival was prolonged for only 2-3 months. Castration Resistant (CR) progression involves variable combinations of clonal selection, ligand-independent androgen receptor (AR) activation and alternative growth factor pathways and dative up-regulation of anti-apoptotic genes. Additional therapeutic strategies targeting molecular mechanisms mediating resistance must be developed.

One strategy to improve therapies in advanced PC involves targeting genes that are activated by androgen withdrawal, either to delay or prevent the emergence of the CR phenotype. Recently, we identified Hsp27 as a highly over-expressed gene in CRPC.

Hsp27 knockdown using antisens oligonucleotides (ASO) and small interference RNA (siRNA) increased apoptotic rates and enhanced hormone- and chemo- therapy in PC.We developed and patented a second generation ASO targeting Hsp27 that has been licensed (OGX-427) and clinical trials phase II is currently in process in PC (http://oncogenex.ca/). Despite OGX-427 efficiency, the functional role of stress induced Hsp27 in castration or chemotherapy-induced apoptosis remains undefined.

Our purpose now is to elucidate the pathways leading to Hsp27 action in CRPC in order to 1/ Increase the pharmacological safety of OGX-427 and obtain the FDA approval 2/find new specific therapeutic targets and treatment strategy for CRPC that would have no toxicity for normal tissues.

Members

Palma Rocchi, CR1 Inserm, Responsable
David Taïeb, MCU-PH, Médecine Nucléaire
Sophie Giusiano-Courcambeck, MCU-PH, Anatomo-Pathologiste
Raquel Mejias-Laguna, Dr. Sciences, Jeune Chercheur, Contrat SATT Sud-Est
Sarah Karaki, Thèse Oncologie, 3ème année, Contrat Amidex
Hajer Ziouziou, Thèse Oncologie, 4ème année, Bourse Alternance Tunisienne
Chayma Cherif, Thèse Oncologie, 1ère année, Bourse Alternance Tunisienne
Tan-Nguyen Dang, Thèse Oncologie, 1ère année, Bourse du Gouv Vietnamien
Nicolas Branger, Interne en Urologie, M2 Onco, Bourse de l'Ass Franç d'Urologie
Aurélien Archier, Assistant en Médecine Nucléaire, M2 Oncologie
Mona Ouled Dhaou, M2 Oncologie, Gratification

Selected publications

Fundings

  • Financement Inserm Transfert « Proof of Concept of Projects with high economical Potential ».
  • Financement Amidex. Novel UltraSonic Biomarkers of Tumor response
  • Financement cancéropole PACA*. Nanodevice for targeted imaging in prostate cancer.
  • Financement Inserm Transfert « Proof of Concept of Projects with high economical Potential »
  • Financement ITMO Cancer « Biologie des Systèmes »
  • Financement CNRS « PEPS : innovation thérapeutique »
  • Financement ANR « Programme Emergence »
  • Financement INCa-ARC-Ligue « Programme d'Action Intégrée sur le cancer de la prostate »
  • Financement Européen « EuroNanoMed »

Hemopathies

The laboratory analyses the molecular alterations of myeloid hemopathies, myelodysplastic and myloproliferative syndromes and chronic myelomonocytic leukemia. We were the first to report mutations of the ASXL1 gene in these hemopathies.

Mutations of ASLX1, together with mutations of RUNX1, constitute a poor prognosis; they are frequent in cases with dysplasia and are characteristic of a particular pathway of leukemogenesis, distinct from that associated with mutations in NPM1/DNMT3A.

We have also identified molecular similarities between refractory anemia with ringed sideroblasts and myelomonocytic leukemia with dysplasia.

Circulating tumor cells & cancer metastases

New theme addressed in the Molecular Oncology department
Head D Birnbaum

Membres :
Alexia Lopresti, PhD Student
Claire Acquaviva, CR1 Inserm
Emilie Mamessier, CR1 Inserm, HDR
Severine Garnier, IR IPC

Summary:
Colorectal cancer (CRC) is one of the most frequent cancers. Despite recent advances in treatment and surgery, the 10-year survival rate is of 50%. As for all cancers, prognosis is largely dependent on the stage at which the cancer is diagnosed and treated. Metastatic spread is the main mortality cause, and in CRC, liver is the main site of metastasis. About 50% of CRC patients develop liver metastasis (synchronous or metachronous liver metastasis, mCCR). Liver resection is the only treatment associated with long-term survival. However, surgery is only possible for 20 to 30% of these patients. For all the other patients, the important challenges remain the early detection and targeting of metastatic malignant cells and the assessment of treatment efficiency. In all these aspects, Circulating Tumor Cells (CTCs) are relevant as they can serve as real-time liquid tumor biopsy. CTCs are believed to detach from the tumor mass, enter the blood circulation and eventually contribute to distant spread in other organs (Figure 1). 

The study of large cohorts of mCCR patients has proved that CTCs detection and abundance are associated with cancer progression, metastases, poor prognosis and risk of relapse (1-3). In this context, CTCs are extremely interesting to study, as models to understand the metastatic process as biomarkers that might guide treatment decisions and as cells to be targeted to avoid metastasis (4,5). However, CTCs represent a heterogeneous population. Only a low percentage has the true potential to grow into solid metastases, whereas most CTCs may just be unable to survive in periphery. Our objective is to characterize such heterogeneity and identify the CTCs with actual metastatic potential. The general lines driving our project are to:

- Compare ex-vivo CTCs (at the single cell level) to the corresponding primary tumor and metastases at the molecular level (using a Biomark/fluidigm approach for transcriptomic analyses and the next generation sequencing for the identification of “druggable” targets by high throughput sequencing),

- Identify new surface CTCs markers (educated guess or issued from the previous analyses), in regards to their metastatic potential, and in the prospect of a specific targeting of these cells (Figure 2),

- Develop an in vitro ecosystem reconstituting interactions between CTCs and their host site. Using 3D liver organoids as the metastatic niche, we would like to build a model predicting which CTCs are more susceptible to seed metastases. Grown CTCs will further be characterized at the molecular levels and compared to the respective patient’s metastases and corresponding xenograft (if available).

Altogether, these approaches will help decipher the mechanisms involved in metastases occurrence and identified adapted targeted therapies, including some that might be useful early during the metastatic process. More specifically, we aim at building a predictive model gathering all the collected information (CTCs counts, phenotype, cluster, mutations, growth and stem cell potential, response to drugs…) able to anticipate metastases occurrences. In the meantime, our approaches will allow the rapid identification of CTCs and a short-term culture to characterize CTCs with a metastatic potential in a time scale compatible with personalized medicine strategies. This project started in 2015. Since, we have launched CTCs description, live CTCs isolation, organotypic cultures and molecular analyses on a first cohort of cancer patients (PERMED study). Shortly, we will be involved in translational projects, such as the follow-up of different cohorts of CRC patients +/- hepatic metastases, in collaboration with clinicians. Other types of cancer will be studied.



1. Ramirez JM, Fehm T, Orsini M, Cayrefourcq L, Maudelonde T, Pantel K, Alix-Panabieres C. Prognostic relevance of viable circulating tumor cells detected by EPISPOT in metastatic breast cancer patients. Clin Chem2014 Jan;60(1):214-21.
2. Deneve E, et al. Capture of viable circulating tumor cells in the liver of colorectal cancer patients. Clin Chem2013 Sep;59(9):1384-92.
3. Bidard FC, et al. Clinical validity of circulating tumour cells in patients with metastatic breast cancer: a pooled analysis of individual patient data. Lancet Oncol2014 Apr;15(4):406-14.
4. Pantel K, Speicher MR. The biology of circulating tumor cells. Oncogene2015 Jun 8.
5. Alix-Panabieres C, Pantel K. Challenges in circulating tumour cell research. Nat Rev Cancer2014 Sep;14(9):623-31.