Diagnostic methods for neural disorders
The invention generally relates to optical methods for the diagnosis of neuronal condition by converting a cell from a patient into a neuron and optically evaluating action potentials of that cell in vitro. The cell is transformed with an optical reporter and exhibits an optical signature in response to neural stimulation. Using genome-editing, a control cell can be made that is isogenic but-for a known mutation and a control signature obtained from the control cell. Thus, methods of the invention reveal potential neurodegenerative effects of a mutation as manifested in a patient's genetic context. The optical signature of the cell, or the difference between the signature and the control signature, is correlated to a diagnosis of the neurodegenerative disease.
1. A method of characterizing a neurological condition, the method comprising:
obtaining a human-derived cell having a mutation associated with a condition;
expressing an exogenous optical reporter and an exogenous optical voltage actuator in the cell;
illuminating the optical voltage actuator expressed by the cell or a neighboring cell, wherein the optical voltage actuator initiates an action potential in response to the illumination as an optical stimulation;
detecting an optical signature generated by the optical reporter in response to the optical stimulation of the cell; and
comparing the detected signature to an expected signature detected from control cell, wherein a difference between the observed signature and the expected signature provides information about the condition.
2. The method of claim 1 , wherein the optical reporter of electrical activity is an exogenous fluorescent protein.
3. The method of claim 2 , wherein the protein comprises at least a portion of a microbial rhodopsin.
4. The method of claim 3 , wherein the protein includes the microbial rhodopsin operably linked to at least one additional fluorescent protein or chromophore.
5. The method of claim 1 , further comprising editing a genome of the cell to include a mutation relative to a control genome.
6. The method of claim 1 , further comprising obtaining the expected signature by obtaining a control cell suspected of not having the condition and observing a control signature generated by a control optical reporter in the control cell.
7. The method of claim 1 , wherein obtaining the cell comprises differentiating a stem cell into a neuron or converting a somatic cell to a neuron.
8. The method of claim 7 , wherein converting the somatic cell into the neuron comprises one selected from the list consisting of: direct lineage conversion; and conversion through an iPS intermediary.
9. The method of claim 1 , wherein the condition comprises one selected from the list consisting of Cockayne syndrome, Down Syndrome, Dravet syndrome, familial dysautonomia, Fragile X Syndrome, Friedreich's ataxia, Gaucher disease, hereditary spastic paraplegias, Machado-Joseph disease, Phelan-McDermid syndrome (PMDS), polyglutamine (polyQ)-encoding CAG repeats, spinal muscular atrophy, Timothy syndrome, Alzheimer's disease, frontotemporal lobar degeneration, Huntington's disease, multiple sclerosis, Parkinson's disease, spinal and bulbar muscular atrophy, and amyotrophic lateral sclerosis.
10. The method of claim 1 , wherein the difference between the observed signature and the expected signal comprises a modified probability of a voltage spike in response to the stimulation of the cell.
11. A method of diagnosing a condition, the method comprising:
obtaining a cell from a patient suspected of having the condition;
expressing an exogenous optical reporter and an exogenous optical voltage actuator in the cell;
illuminating the optical voltage actuator expressed by the cell or a neighboring cell, wherein the optical voltage actuator initiates an action potential in response to the illumination as an optical stimulation;
detecting an optical signature from the optical reporter in response to the optical stimulation of the cell; and
comparing the detected signature to an expected signature detected from control cell, wherein a difference between the observed signature and the expected signature corresponds to a diagnosis of the condition; and
generating a report that includes the diagnosis for the patient.
12. The method of claim 11 , further comprising editing a genome of the cell to include a mutation relative to a control genome.
13. The method of claim 11 , further comprising obtaining the expected signature by obtaining a control cell suspected of not having the condition and observing a control signature generated by a control optical reporter in the control cell.
14. The method of claim 13 , wherein the control cell and the cell are isogenic but for a mutation.
15. The method of claim 11 , wherein obtaining the cell comprises one selected from the group consisting of:
differentiating a stem cell into a neuron; and
direct lineage conversion of a somatic cell into a neuron.
16. The method of claim 11 , wherein detecting an optical signature comprises detecting a cluster of different cells with a microscope and using a computer to isolate the signature generated by the optical reporter from a plurality of signature from the different cells.
17. The method of claim 11 , wherein the condition comprises one selected from the list consisting of Cockayne syndrome, Down Syndrome, Dravet syndrome, familial dysautonomia, Fragile X Syndrome, Friedreich's ataxia, Gaucher disease, hereditary spastic paraplegias, Machado-Joseph disease, Phelan-McDermid syndrome (PMDS), polyglutamine (polyQ)-encoding CAG repeats, spinal muscular atrophy, Timothy syndrome, Alzheimer's disease, frontotemporal lobar degeneration, Huntington's disease, multiple sclerosis, Parkinson's disease, and spinal, bulbar muscular atrophy, and amyotrophic lateral sclerosis.
18. The method of claim 11 , wherein the difference between the observed signature and the expected signature comprises a modified probability of a voltage spike in response to the stimulation of the cell relative to a control.