ch has further identifi ed Ahi 1/AHI 1 as a novel modulator of BCR ABL that . Identifi cation of an AHI 1 BCR ABL JAK2 interaction complex raises many interesting questions about Panobinostat HDAC inhibitor the nature of the interaction, how it is regulated, and how it contributes to malignant transformation, altered BCR ABL signaling, and IM response/resistance of CML stem/progenitor cells. Particularly notable is the observation that coexpression of Ahi 1 in BCR ABL inducible cells can rescue GF independent cell growth that is inhibited by down regulation of BCR ABL. Interestingly, this renewed GF independence with introduction of Ahi 1 appears to be regulated by sustained phosphorylation of BCR ABL, rather than its continual expression, as these eff ects were observed in cotransduced cells where BCRABL expression was inducibly suppressed in vitro.
These results suggest that physical interaction between Ahi 1 and BCR ABL may stabilize a protein protein interaction complex that enables constitutively active BCR ABL tyrosine kinase activity and further alters specifi c downstream BCR ABL signaling pathways that deregulate cellular proliferation and apoptosis control. This is further supported BIX 02189 by identifi cation of JAK2 as an associated protein in this interaction complex and observation of enhanced activity of the JAK2 STAT5 pathway in BCR ABL inducible cells with cotransduction of Ahi 1 in the presence of GF stimulation with IL 3. Interestingly, all these fi ndings can be replicated signifi cantly in a human CML cell line system in which changes in AHI 1 expression are found to modulate tyrosine phosphorylation and protein expression of BCR ABL and JAK2 STAT5.
It is known that BCRABL signaling closely mimics signaling pathways of cytokine receptors and that both IL 3/GM CSF receptor activation and the BCR ABL oncoprotein can induce a tyrosine phosphorylation cascade of which numerous proteins, including JAK2 and STAT5, are common substrates. Interestingly, BCR ABL expressing cells have many similarities to cells induced by IL 3 stimulation or cells with forced IL 3 overexpression. Early studies have shown that BCR ABL can interact with the common chain of IL 3/GM CSF receptor and constitutively activate JAK2.
In particular, increased phosphorylation of STAT5, which was previously thought to be an immediate function of the BCR ABL oncoprotein, has now been shown in primary CML CD34 progenitor cells to occur largely as a consequence of BCRABL induced activation of IL 3 autostimulation, leading to activation of STAT5. It has been reported that targeted disruption of the STAT5A and STAT5B genes reduces myeloid progenitor numbers, suggesting a nonredundant role for STAT5 in primitive normal hematopoiesis. Recent studies further suggest that autocrine production of GM CSF may contribute to IM and NL resistance in BCR ABL progenitors through activation of JAK2 and STAT5 pathway, and inhibition of STAT5 expression by a shRNA approach signifi cantly reduced colony formation of CD34 CML progenitor cells in vitro. In addition, JAK2 is known to interact with the C terminal region of BCR ABL, and recent studies further show that mouse hematopoietic cells transformed by the T315I mutant of BCR ABL can be induced to undergo apoptos