Organotypic three-dimensional patient derived metastatic colorectal cancer model to mimic in vitro the metastases biology - Workshop Termis

Organotypic three-dimensional patient derived metastatic colorectal
            cancer model to mimic in vitro the metastases biology
               Francesca Sensi1,5 ,Edoardo D’Angelo1,2, Dipa Natarajan3,4, Matteo Fassan6, Enzo Mammano7, Pierluigi Pilati7,
                  Piero Pavan1,8, Silvia Bresolin1, Gaya Spolverato2, Martina Piccoli1,9, Luca Urbani3,4* and Marco Agostini1,2*.

1   Fondazione Istituto di Ricerca Pediatrica Città della Speranza, Padua, Italy 2 Department of Surgery, Oncology and Gastroenterology, University of Padova, Italy3
                Institute of Hepatology, Foundation for Liver Research, London, UK 4 Faculty of Life Sciences & Medicine, King’s College London, UK
                  5 Department of Women’s and Children’s Health, University of Padova, Italy.6 Department of Medicine, University of Padova, Italy.
                       7 Unit of Surgical Oncology of the Esophagus and Digestive Tract, Veneto Institute of Oncology IOV-IRCCS, Padua, Italy.
                8 Department of Industrial Engineering, University of Padova, Italy. 9 Department of Biomedical Sciences, University of Padova, Italy.

Colorectal cancer liver metastasis (CRLM): facts

• Liver is the most common site of CRC metastasis
  due to its anatomical position regarding its portal
  circulation;

• At the time of diagnosis, up to 25% of patients
  have a synchronous liver metastasis.
  After resection of the primary tumor, approximately
  45% of the patients will diagnosed with liver
  metastasis within the next 3 years;

• 5-year survival rate after curative surgery is
  25-44%

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Aims of the study

                                                                      Piccoli et al. 2018

                      To develop an in vitro patient-derived
                      3D model of CRLM that can mimic in
                           vitro the patient's disease:

• To recapitulate CRLM structural and biochemical microenvironment;

• To evaluate behaviour of cells in progression and metastases: proliferation,
  migration, ECM remodelling;

• To establish a pre-clinical tool for anticancer drug screening assay.

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Decellularized tissue evaluation

                                                                                   pre   post
                                                                                   DET   DET
                                                                 Phenotypical
                                                                 appearance

                                                                 Cellularization

                                                                 ECM
                                                                 components
                                                                 Structure and
                                                                 architecture

                                                                 Ultrastructure

                                            Scale bar = 100 µm

  Decellularization of CRLM tissues produce scaffolds that recapitulate
histological, structural and ultrastructural properties of the native tissue

                                      3/8

Organotypic effects of dECM on…

                              Proliferation

                                               HC   CRC     HL CRLM

                                                        Scale bar = 100 µm

CRC scaffold supports a more proliferative phenotype compared to the
       respective healthy counterpart and to CRLM scaffold

Crucial role of the acellular scaffolds in conditioning tumor cell behavior

                                    4/8

Organotypic effects of dECM on…

                     Migration

                                 Scale bar = 100 µm

CRLM promotes more migration compared to the respective
   healthy counterpart and compared to CRC scaffold

                        5/8

Gene expression profile analysis

                                           Gene Ontology   GSEA Hallmark dataset
                                2D

                                3D CRLM

  These results suggest a
    reduced uncontrolled
 proliferation in 3D cultured
  cells, possibly in favor of
 other pathways to adapt to
the surrounding environment

                                     6/8

5-FU response: 2D vs 3D

   HT-29 cells grown in CRLM scaffolds and exposed to conventional
       IC50 concentration of 5-FU, are resistant to the treatment;
     HT-29 cells grown in 2D significantly reduce proliferation rate.

Therapeutic response to treatment of CRC cells is affected by the complex
         biological microenvironment in which they are cultured

                                   7/8

Conclusions

i) decellularization of CRLM tissues produces scaffolds that recapitulate histological,
structural and ultrastructural properties of the native tissue;

 ii) CRLM scaffolds support CRC cells adhesion and influence proliferation and migration in a
tissue-specific manner;

iii) tissue-specific scaffolds stimulate different gene expression patterns in tumor cells
cultured in 3D environment compared to 2D;

iv) CRC cells respond differently to standard chemotherapy when cultured in organotypic
scaffolds compared to 2D conditions.

              3D patient-derived model better recapitulates the metastatic
              microenvironment compared to 2D culture and represents a
                 relevant approach for the study of liver CRC metastasis
                               formation and progression

                                            8/8

Thanks to…

  Istituto di Ricerca Pediatrica   Dept. of Surgical, Oncological and     King’s College of London
Fondazione Città della Speranza     Gastroenterological Sciences;       Foundation for Liver Research
     Tissue Engineering lab                 Dept. Of Medicine             Liver Regeneration Group

      Dr.ssa Martina Piccoli             Dr. Edoardo D’Angelo                  Dr. Luca Urbani
        Prof. Piero Pavan                  Dr. Marco Agostini               Dr.ssa Dipa Natarajan
                                        Prof. Salvatore Pucciarelli          Prof. Roger Williams
                                          Prof. Matteo Fassan
                                          Prof. Massimo Rugge