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Peripheral blood cells from children with RASopathies show enhanced spontaneous colonies growth in vitro and hyperactive RAS signaling.

Gaipa G, Bugarin C, Cianci P, Sarno J, Bonaccorso P, Biondi A, Selicorni A - Blood Cancer J (2015)

Bottom Line: Higher spontaneous growth of both burst-forming units-erythroid (BFU-E) and CFU-granulocyte/macrophage (CFU-GM) colonies from RAS-mutated patients were observed as compared with control subjects.We also observed a significantly higher amount of GM-colony-stimulating factor-induced p-ERK in children with RASopathies.Collectively, our data provide a basis for further investigating of only partially characterized hematological alterations present in children suffering from RASopathies, and may provide new markers for progression toward malignant MPD in these patients.

View Article: PubMed Central - PubMed

Affiliation: M. Tettamanti Research Centre, Pediatric Clinic, University of Milano-Bicocca, Monza, Italy.

ABSTRACT
Germline mutations in genes coding for molecules involved in the RAS/RAF/MEK/ERK pathway are the hallmarks of a newly classified family of autosomal dominant syndromes termed RASopathies. Myeloproliferative disorders (MPDs), in particular, juvenile myelomonocytic leukemia, can lead to potentially severe complications in children with Noonan syndrome (NS). We studied 27 children with NS or other RASopathies and 35 age-matched children as control subjects. Peripheral blood (PB) cells from these patients were studied for in vitro colony-forming units (CFUs) activity, as well as for intracellular phosphosignaling. Higher spontaneous growth of both burst-forming units-erythroid (BFU-E) and CFU-granulocyte/macrophage (CFU-GM) colonies from RAS-mutated patients were observed as compared with control subjects. We also observed a significantly higher amount of GM-colony-stimulating factor-induced p-ERK in children with RASopathies. Our findings demonstrate for the first time that PB cells isolated from children suffering from NS or other RASopathies without MPD display enhanced BFU-E and CFU-GM colony formation in vitro. The biological significance of these findings clearly awaits further studies. Collectively, our data provide a basis for further investigating of only partially characterized hematological alterations present in children suffering from RASopathies, and may provide new markers for progression toward malignant MPD in these patients.

No MeSH data available.


Related in: MedlinePlus

Representative quantitative BFU-E (a) and CFU-GM (b) colony numbers as assessed by clonogenic assays. t test, ***P<0.001.
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fig2: Representative quantitative BFU-E (a) and CFU-GM (b) colony numbers as assessed by clonogenic assays. t test, ***P<0.001.

Mentions: Clonogenic assays from PB precursor cells showed a significantly higher spontaneous growth rate of both BFU-E and CFU-GM in RAS-mutated patients (n=22) than control subjects (n=22). We observed an increased number of BFU-E colonies (mean±s.e.m.): 84.45±7.37% vs 30.50±4.32%, P<0.0001, as well as enhanced growth of CFU-GM, (21.14±3.15% vs 7.23±1.38%, P=0.0002; Figures 2a and b, respectively). Augmented colony formation activity of RAS-mutated cells was also observed upon incubation in growth factor-supplemented medium: BFU-E (78.41±6.60% vs 31.14±4.80%, P<0.0001) and CFU-GM (21.59±2.93% vs 8.95±1.77%, P=0.0006).


Peripheral blood cells from children with RASopathies show enhanced spontaneous colonies growth in vitro and hyperactive RAS signaling.

Gaipa G, Bugarin C, Cianci P, Sarno J, Bonaccorso P, Biondi A, Selicorni A - Blood Cancer J (2015)

Representative quantitative BFU-E (a) and CFU-GM (b) colony numbers as assessed by clonogenic assays. t test, ***P<0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526778&req=5

fig2: Representative quantitative BFU-E (a) and CFU-GM (b) colony numbers as assessed by clonogenic assays. t test, ***P<0.001.
Mentions: Clonogenic assays from PB precursor cells showed a significantly higher spontaneous growth rate of both BFU-E and CFU-GM in RAS-mutated patients (n=22) than control subjects (n=22). We observed an increased number of BFU-E colonies (mean±s.e.m.): 84.45±7.37% vs 30.50±4.32%, P<0.0001, as well as enhanced growth of CFU-GM, (21.14±3.15% vs 7.23±1.38%, P=0.0002; Figures 2a and b, respectively). Augmented colony formation activity of RAS-mutated cells was also observed upon incubation in growth factor-supplemented medium: BFU-E (78.41±6.60% vs 31.14±4.80%, P<0.0001) and CFU-GM (21.59±2.93% vs 8.95±1.77%, P=0.0006).

Bottom Line: Higher spontaneous growth of both burst-forming units-erythroid (BFU-E) and CFU-granulocyte/macrophage (CFU-GM) colonies from RAS-mutated patients were observed as compared with control subjects.We also observed a significantly higher amount of GM-colony-stimulating factor-induced p-ERK in children with RASopathies.Collectively, our data provide a basis for further investigating of only partially characterized hematological alterations present in children suffering from RASopathies, and may provide new markers for progression toward malignant MPD in these patients.

View Article: PubMed Central - PubMed

Affiliation: M. Tettamanti Research Centre, Pediatric Clinic, University of Milano-Bicocca, Monza, Italy.

ABSTRACT
Germline mutations in genes coding for molecules involved in the RAS/RAF/MEK/ERK pathway are the hallmarks of a newly classified family of autosomal dominant syndromes termed RASopathies. Myeloproliferative disorders (MPDs), in particular, juvenile myelomonocytic leukemia, can lead to potentially severe complications in children with Noonan syndrome (NS). We studied 27 children with NS or other RASopathies and 35 age-matched children as control subjects. Peripheral blood (PB) cells from these patients were studied for in vitro colony-forming units (CFUs) activity, as well as for intracellular phosphosignaling. Higher spontaneous growth of both burst-forming units-erythroid (BFU-E) and CFU-granulocyte/macrophage (CFU-GM) colonies from RAS-mutated patients were observed as compared with control subjects. We also observed a significantly higher amount of GM-colony-stimulating factor-induced p-ERK in children with RASopathies. Our findings demonstrate for the first time that PB cells isolated from children suffering from NS or other RASopathies without MPD display enhanced BFU-E and CFU-GM colony formation in vitro. The biological significance of these findings clearly awaits further studies. Collectively, our data provide a basis for further investigating of only partially characterized hematological alterations present in children suffering from RASopathies, and may provide new markers for progression toward malignant MPD in these patients.

No MeSH data available.


Related in: MedlinePlus