1. Bando Ricerca Finalizzata 2009 – Ministero della Salute
Progetto Giovani Ricercatori
Fibrociti e cellule mesenchimali staminali nella fibrosi polmonare idiopatica
Stefania Cerri, Paolo Spagnolo, Fabrizio Luppi, Luca Richeldi
Meeting DAI di Oncologia, Ematologia e Patologie dell’Apparato Respiratorio
Gaiato, 16 luglio 2011

2. Fibrocytes UNIVERSITY OF MODENA AND REGGIO EMILIA
Firstly described in 19941
Circulating, bone marrow-derived cells with the ability to adopt a mesenchymal phenotype
Express a combination of hematopoietic and mesenchymal markers
In normal individuals, approx. 0.5% of circulating leukocytes
UNIVERSITY OF MODENA
AND REGGIO EMILIA
Control
Collagen I
1. Bucala R., et al. Molecular Medicine, 1994; 1(1):71-81
SINCE 1175

3. Fibrocytes markers UNIVERSITY OF MODENA AND REGGIO EMILIA
They are known to be positive for both CD45 and Collagen I or pro-Collagen I
They may express CD34 (stem cell marker)
According to some authors, they also share monocyte markers (such CD14 and CD11)
Chemokines receptors (such as CXCR4)
Lack of expression of CD3, CD4, CD8, CD19 and CD25
They seem to be able to present antigens
UNIVERSITY OF MODENA
AND REGGIO EMILIA
SINCE 1175

4. Fibrocytes markers UNIVERSITY OF MODENA AND REGGIO EMILIA
Bellini A., Mattioli S., Laboratory Investigation 2007; 87:858-70
SINCE 1175

5. Fibrocytes in health and disease
Fibrocytes have been described in several diseases in which fibrosis occurs
Asthma
Skin scars and keloids
Scleroderma
Liver fibrosis
Cardiac disease
Renal fibrosis
Circulating fibrocytes may be increased in normal aging
UNIVERSITY OF MODENA
AND REGGIO EMILIA
Herzog EL., Bucala R., Experimental Hematology 2010; 38:548-56
SINCE 1175

6. IDIOPATHIC PULMONARY FIBROSIS (IPF)
Specific form of chronic, progressive fibrosing interstitial pneumonia of unknown cause, occurring in adults and limited to the lungs.
UNIVERSITY OF MODENA
AND REGGIO EMILIA
ATS/ERS/JRS/ALAT IPF GUIDELINES 2011
SINCE 1175

7. UNIVERSITY OF MODENA
AND REGGIO EMILIA
SINCE 1175

8. Pathogenesis of IPF Abnormal wound healing model
UNIVERSITY OF MODENA
AND REGGIO EMILIA
Selman M., Ann Intern Med 2001; 134:136.
SINCE 1175

9. UNIVERSITY OF MODENA
AND REGGIO EMILIA
SINCE 1175

10. Circulating fibrocytes in human fibrotic interstitial lung disease
UNIVERSITY OF MODENA
AND REGGIO EMILIA
UIP: usual interstitial pneumonia; NSIP: non-specific interstitial pneumonia
Mehrad et al. Biochem Biophys Res Commun 2007; 353:104-8.
SINCE 1175

11. Circulating fibrocytes and IPF prognosis
UNIVERSITY OF MODENA
AND REGGIO EMILIA
Fibrocytes defined as CD45+ and Coll-1+ cells among live PBMC
Moeller et al. Am J Respir Crit Care Med 2009; 179:
SINCE 1175

12. Circulating fibrocytes and IPF prognosis
UNIVERSITY OF MODENA
AND REGGIO EMILIA
Fibrocytes defined as CD45+ and Coll-1+ cells among live PBMC
Moeller et al. Am J Respir Crit Care Med 2009; 179:
SINCE 1175

13. AGE MATTERS
Ageing mice develop more fibrosis in response to Bleomycin, have more fibrocytes
Xu J. et al. J Gerontol A Biol Sci Med Sci 2009
Figure 6. BMDMSCs obtained from bone marrow of SAMP8 mice had a decreased ability to differentiate into chondrocytes. To demonstrate if the potential of BMDMSCs to differentiate is modified with age, BMDMSCs obtained from 12-month-old SAMP8 and SAMR1 mice were cultured in a chondrogenic basal media and adipogenic media for 14 days. Transformed cells were visualized by staining the cells: alcian blue for chondrocytes and oil red O staining for adipocytes. (A) Positive transformed cells stained blue (arrow) under light microscopy, 20x and 100x magnification. (B) The number of transformed cells was determined by counting the number of positive of cells for every 100 nucleus. Notes: BMDMSC = bone marrow–derived stem cell; SAMP = senescence-accelerated prone mice; SAMR = senescence-accelerated resistant mice. n = 4. *p < .01.
MSCs of ageing mice have decreased ability to differentiate

14. Mesenchymal stem cell engraftment in lung is enhanced in response to bleomycin exposure and ameliorates its fibrotic effects
Saline Bleomycin Bleomycin + BM-MSC
Ortiz LA et al. PNAS 2003; 100(14):

15. IL PROGETTO
UNIVERSITY OF MODENA
AND REGGIO EMILIA
MESENCHYMAL PROGENITOR CELLS AS BIOMARKERS IN IDIOPATHIC PULMONARY FIBROSIS: A PROSPECTIVE STUDY GR
SINCE 1175

16. Project aims
Aim 1
Measurement and characterization of circulating fibrocytes
Longitudinal measurement of circulating fibrocytes in IPF patients and comparison with clinical outcomes
Characterization of expression markers related to lung homing and differentiation
Assessment of cytokine levels in the serum of IPF patients
Evaluation of the effects of cytokine stimulation on in vitro proliferation, differentiation and migration ability of fibrocytes isolated from IPF patients.

17. Project aims Aim 2 Measurement and characterization of MSC
Characterization of bone-marrow derived MSC from IPF patients (numbers and expression markers)
Correlation between MCS and fibrocytes numbers
In vitro assessment of differentiation potential of MSC derived from IPF patients.

18. Study design UNIVERSITY OF MODENA AND REGGIO EMILIA
Total duration: 3 years
Patients with a diagnosis of IPF according to ATS/ERS/JRS Guidelines 2011
Prospective enrollment of both prevalent and incident cases
Collection of blood samples for fibrocytes and cytokine measurement at the enrollment and then every 6 months during the follow-up
Bone marrow samples
UNIVERSITY OF MODENA
AND REGGIO EMILIA
SINCE 1175

19. with precious feedback and suggestions from
“Working group”
Stefania Cerri
Fabrizio Luppi
Paolo Spagnolo
Francesca Panico
Luca Richeldi
with precious feedback and suggestions from
Giulia Grisendi
Massimo Dominici
UNIVERSITY OF MODENA
AND REGGIO EMILIA
SINCE 1175