Method for diagnosing a protein misfolding disease using nerve cells derived from iPS cells
The present invention provides a method for detecting onset of, or the risk of development of, a protein misfolding disease, and a method for predicting the age of onset of a protein misfolding disease using nerve cells derived from iPS cells. The present invention also provides a kit to be used in these methods.
1. A method for diagnosing whether a test subject has developed Alzheimer's disease, wherein the test subject has a risk of developing Alzheimer's disease or is suspected of having Alzheimer's disease, said method comprising the steps of:
(a) establishing iPS cells from somatic cells derived from the test subject;
(b) inducing differentiation of said iPS cells into nerve cells;
(c) measuring the amount of a causative protein selected from the amount of Aβ40, the amount of Aβ42, or the ratio of the amount of Aβ42 to that of Aβ40, or the activity or the expression level of an enzyme involved in degradation of the causative protein in said nerve cells, wherein the enzyme is neprilysin;
(d) comparing said measured value with the amount of the causative protein in control cells, or with the activity or the expression level of the enzyme involved in degradation of the causative protein in control cells, wherein said control cells are nerve cells obtained by inducing differentiation of iPS cells produced from somatic cells derived from a control subject who has not developed Alzheimer's disease at the same age as the test subject, and when the amount of said causative protein in the test subject is higher than that in the control subject, or when the activity or the expression level of neprilysin in the test subject is lower than that in the control subject, it is determined that the test subject has developed Alzheimer's; and
(e) determining if the subject has developed Alzheimer's disease based upon the comparison in step (d).
2. The method according to claim 1 , wherein said subject is human.
3. The method according to claim 1 , wherein said differentiation is performed by culturing with media contained B27-supplement and N2-supplement.
4. A method for diagnosing whether a test subject has developed Alzheimer's disease, wherein the test subject has a risk of developing Alzheimer's disease or is suspected of having Alzheimer's disease, said method comprising the steps of:
(a) establishing iPS cells from somatic cells derived from the test subject;
(b) inducing differentiation of said iPS cells into nerve cells;
(c) measuring the amount of a causative protein selected from the amount of Aβ40, the amount of Aβ42, or the ratio of the amount of Aβ42 to that of Aβ40, or the activity or the expression level of an enzyme involved in degradation of the causative protein in said nerve cells, wherein the enzyme is neprilysin;
(d) comparing said measured value with the amount of the causative protein in control cells, or with the activity or the expression level of the enzyme involved in degradation of the causative protein in control cells, wherein said control cells are nerve cells obtained by inducing differentiation of iPS cells produced from somatic cells derived from a control subject who has already developed Alzheimer's disease at the same age as the test subject, and when the amount of said causative protein in the test subject is higher than or equivalent to that in the control subject, or when the activity or the expression level of neprilysin in the test subject is lower than or equivalent to that in the control subject, it is determined that the test subject has developed Alzheimer's disease; and
(e) determining if the subject has developed Alzheimer's disease based upon the comparison in step (d).
5. The method according to claim 4 , wherein said subject is human.
6. The method according to claim 4 , wherein said differentiation is performed by culturing with media contained B27-supplement and N2-supplement.