Eric Klann, PhD, Principal Investigator (2003-present)
Aditi Bhattacharyya, PhD, FRAXA Postdoctoral Fellow (2011-2012)
Lingfei Hou, PhD, FRAXA Postdoctoral Fellow (2008-2010)

FRAXA Awards:

Aditi Bhattacharya, Eric Klann, 9/20/2012

by Eric Klann, 6/8/2011

It is well known that fragile X syndrome (FXS) causes exaggerated rates of protein synthesis (translation), due to the lack of fragile X mental retardation protein (FMRP), a suppressor of translation. mTORC1 is critical for the initiation of translation, which it controls by phosphorylating several substrates, including p70 S6 kinase1 (S6K1).

In previous studies conducted in collaboration with Dr. Suzanne Zukin's laboraotory at Albert Einstein School of Medicine, we found that FXS model mice exhibit an upregulation in mTOR-dependent translational control, including an increase in S6K1 phosphorylation. These findings led us to hypothesize that genetic manipulations that remove S6K1 would reverse increased phosphorylation of S6K1 substrates, exaggerated protein synthesis, aberrant neuronal morphology, altered synaptic plasticity, and abnormal behaviors in FXS model mice. In previous studies funded by FRAXA, we have found that genetic ablation of S6K1 prevents many of the aforementioned abnormalities displayed by FXS model mice.

Our findings with genetic reduction of S6K1, though extremely exciting, are not likely to be of utility as a treatment option for individuals with FXS. Thus, identifying chemical agents that achieve inhibition of S6K1 would likely be of more use as a therapy for FXS. Our currently funded FRAXA project aims to assess the efficacy of pharmacologically targeting of S6K1 to reverse phenotypes observed in FXS model mice using a novel, selective inhibitor of S6K1. We will conduct a series experiments with the S6K1 inhibitor to determine whether it can reverse the molecular, morphological, synaptic, and behavioral abnormalities displayed by FXS model mice. Findings from our studies will provide important information about the viability of S6K1 as a novel therapeutic target for treatment of FXS.

By Eric Klann, 7/1/2008

The mGluR theory of fragile X syndrome (FXS) states that the functional consequences of mGluR-dependent protein synthesis are exaggerated when the fragile X protein (FMRP) is absent. Our previous studies funded by FRAXA are consistent with this theory in that it appears that the baseline levels of proteins that are synthesized (translated from mRNA) in response to mGluR stimulation during mGluR-LTD are elevated in FXS model mice. However, our data also suggest that although there is exaggerated basal translation of these mRNAs into protein, there is no further mGluR-dependent protein synthesis during mGluR-dependent long-term depression (mGluR-LTD) in the FXS model mice.

mTOR is a protein kinase that is critical for the initiation of translation, which it controls by phosphorylating several substrates, including p70 S6 kinase1 (S6K1). As mentioned above, we have found that baseline levels of proteins that are synthesized in response to mGluR stimulation during mGluR-LTD are elevated in FXS model mice, and that there is no further mGluR-dependent synthesis of these proteins during mGluR-LTD in the FXS model mice. These findings suggest that there may be an upregulation of mTOR-dependent translational control pathways in FXS model mice. Consistent with this idea, in preliminary studies conducted in collaboration with Ali Sharma and Suzanne Zukin at Albert Einstein School of Medicine we have found that FXS model mice exhibit an upregulation in mTOR-dependent translational control, including an increase in S6K1 phosphorylation.

The preliminary results described above have provided important information concerning the elevation of basal translational control in FXS model mice. These results have led us to hypothesize that genetic manipulations that reduce the elevated basal levels of S6K1 phosphorylation in the FXS model mice will reverse aberrant neuronal morphology, enhanced mGluR-LTD, and abnormal behaviors observed in these mice. These experiments will test several aspects of the mGluR theory of FXS put forward by Mark Bear and colleagues, and will provide critical information concerning the therapeutic potential of S6K1 antagonists for treatment of patients with FXS.

Summary of Previous Work

In our first series of studies funded by FRAXA, we identified several signaling pathways that couple mGluRs to the protein translation machinery during metabotropic glutamate receptor-dependent long-term depression (mGluR-LTD). These pathways pointed to potential mRNAs that could be rapidly translated after the induction of LTD. We observed that rapid translation of several mRNAs occurs during LTD in normal mice, and that translation of these mRNAs, including MAP1b, was misregulated in mice that model fragile X syndrome (FXS). In complementary studies, we studied mGluR-LTD in mice that overexpress human FMRP (YAC FMR1 transgenic mice) and found that LTD was inhibited in the YAC FMR1 transgenic mice. Finally, we found that mGluR-LTD was not only associated with the rapid translation of FMRP, but also a rapid degradation of FMRP that was required for mGluR-LTD. These findings were published in Neuron in August 2006 (Neuron 51: 441-454).

As mentioned above, we identified several proteins whose levels are rapidly increased during mGluR-LTD in wild-type mice. In each case, the baseline levels of these proteins were increased in the brains of FXS model mice. Furthermore, we found that mGluR-LTD in FXS mice did not result in an increase in the levels of these proteins. One of these proteins was MAP1b. In a second series of studies funded by FRAXA, we focused on mGluR5-dependent regulation of MAP1b during mGluR-LTD in FXS model mice. We used mGluR5 and MAP1b knockout mice to determine whether the enhanced mGluR-LTD in FXS model mice could be prevented when the levels of mGluR5 and MAP1b were decreased by genetic manipulations by 50%. testing testing testing testing testing testing testing testingt