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Label-free, live optical imaging of reprogrammed bipolar disorder patient-derived cells reveals a functional correlate of lithium responsiveness.

Wang JL, Shamah SM, Sun AX, Waldman ID, Haggarty SJ, Perlis RH - Transl Psychiatry (2014)

Bottom Line: However, efficiently identifying and characterizing relevant neuronal phenotypes in the absence of well-defined pathophysiology remains a challenge.In this study, we collected fibroblast samples from patients with bipolar 1 disorder, characterized by their lithium response (n=12), and healthy control subjects (n=6).We identified a cellular phenotype in reprogrammed neurons using a label-free imaging assay based on a nanostructured photonic crystal biosensor and found that an optical measure of cell adhesion was associated with clinical response to lithium treatment.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Psychiatry, Center for Experimental Drugs and Diagnostics, Massachusetts General Hospital, Boston, MA, USA [2] Center for Human Genetics Research, Massachusetts General Hospital, Boston, MA, USA [3] Stanley Center for Psychiatric Research, Broad Institute of MIT & Harvard, Cambridge, MA, USA.

ABSTRACT
Development of novel treatments and diagnostic tools for psychiatric illness has been hindered by the absence of cellular models of disease. With the advent of cellular reprogramming, it may be possible to recapitulate the disease biology of psychiatric disorders using patient skin cells transdifferentiated to neurons. However, efficiently identifying and characterizing relevant neuronal phenotypes in the absence of well-defined pathophysiology remains a challenge. In this study, we collected fibroblast samples from patients with bipolar 1 disorder, characterized by their lithium response (n=12), and healthy control subjects (n=6). We identified a cellular phenotype in reprogrammed neurons using a label-free imaging assay based on a nanostructured photonic crystal biosensor and found that an optical measure of cell adhesion was associated with clinical response to lithium treatment. This cellular phenotype may represent a useful biomarker to evaluate drug response and screen for novel therapeutics.

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Generation of BD patient-derived iNLCs and BIND imaging. (a) Fibroblasts transduced with miR-9/9*-124, NEUROD2, ASCL1 and MYT1L undergo selection 3 days after lentiviral infection. RFP expression indicates miR-9/9*-124 expression. Morphological changes occur while cells are undergoing selection. At 14 days posttransduction, cells have elongated processes resembling neurite outgrowth. (b) Nanostructured photonic crystal biosensors form the bottom surface of a 384-well plate onto which cells can adhere. Collimated broadband light is projected onto the biosensor, and the resulting resonant reflected light is collected and recorded by an imaging spectrometer, illustrated in c. (d) An overlay of the RFP indicator and BIND scan show that detailed morphology of cells can be captured by the BIND scan and is similar to the morphology seen by fluorescent light microscopy of the RFP indicator. (e) Incubated BIND Scanner allows long-term time-lapse imaging of iNLCs. BD, bipolar disorder; iNLC, induced neuronal-like cell; RFP, red fluorescent protein.
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fig2: Generation of BD patient-derived iNLCs and BIND imaging. (a) Fibroblasts transduced with miR-9/9*-124, NEUROD2, ASCL1 and MYT1L undergo selection 3 days after lentiviral infection. RFP expression indicates miR-9/9*-124 expression. Morphological changes occur while cells are undergoing selection. At 14 days posttransduction, cells have elongated processes resembling neurite outgrowth. (b) Nanostructured photonic crystal biosensors form the bottom surface of a 384-well plate onto which cells can adhere. Collimated broadband light is projected onto the biosensor, and the resulting resonant reflected light is collected and recorded by an imaging spectrometer, illustrated in c. (d) An overlay of the RFP indicator and BIND scan show that detailed morphology of cells can be captured by the BIND scan and is similar to the morphology seen by fluorescent light microscopy of the RFP indicator. (e) Incubated BIND Scanner allows long-term time-lapse imaging of iNLCs. BD, bipolar disorder; iNLC, induced neuronal-like cell; RFP, red fluorescent protein.

Mentions: Skin samples from a clinically phenotyped cohort of patients with bipolar 1 disorder and a history of lithium treatment (n=12) and matched screened healthy control subjects (n=6) were expanded into fibroblast lines and subsequently transformed using recombinant lentiviruses carrying transgenes for miR-9/9*-124, NEUROD2, ASCL1 and MYT1L.9 This transformation results in iNLCs, directly transdifferentiated from the starting fibroblast population. The full duration of time to generate ‘mature' neuronal-like cells that express synaptic markers and fire action potentials as described by Yoo et al.9 is 4–5 weeks. However, due to reduced viability we observed over this prolonged time period of differentiation, and the goal of a more rapid screening process, we elected to measure cellular activity at an earlier time point in the transdifferentiation process to facilitate generating reproducible measurements over larger numbers of iNLCs. This period, 13–14 days posttransduction, occurs after antibiotic selection for cells expressing the transgenes, but before the large amount of cellular attrition. Figure 2a shows the timeline of the neural induction.


Label-free, live optical imaging of reprogrammed bipolar disorder patient-derived cells reveals a functional correlate of lithium responsiveness.

Wang JL, Shamah SM, Sun AX, Waldman ID, Haggarty SJ, Perlis RH - Transl Psychiatry (2014)

Generation of BD patient-derived iNLCs and BIND imaging. (a) Fibroblasts transduced with miR-9/9*-124, NEUROD2, ASCL1 and MYT1L undergo selection 3 days after lentiviral infection. RFP expression indicates miR-9/9*-124 expression. Morphological changes occur while cells are undergoing selection. At 14 days posttransduction, cells have elongated processes resembling neurite outgrowth. (b) Nanostructured photonic crystal biosensors form the bottom surface of a 384-well plate onto which cells can adhere. Collimated broadband light is projected onto the biosensor, and the resulting resonant reflected light is collected and recorded by an imaging spectrometer, illustrated in c. (d) An overlay of the RFP indicator and BIND scan show that detailed morphology of cells can be captured by the BIND scan and is similar to the morphology seen by fluorescent light microscopy of the RFP indicator. (e) Incubated BIND Scanner allows long-term time-lapse imaging of iNLCs. BD, bipolar disorder; iNLC, induced neuronal-like cell; RFP, red fluorescent protein.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150245&req=5

fig2: Generation of BD patient-derived iNLCs and BIND imaging. (a) Fibroblasts transduced with miR-9/9*-124, NEUROD2, ASCL1 and MYT1L undergo selection 3 days after lentiviral infection. RFP expression indicates miR-9/9*-124 expression. Morphological changes occur while cells are undergoing selection. At 14 days posttransduction, cells have elongated processes resembling neurite outgrowth. (b) Nanostructured photonic crystal biosensors form the bottom surface of a 384-well plate onto which cells can adhere. Collimated broadband light is projected onto the biosensor, and the resulting resonant reflected light is collected and recorded by an imaging spectrometer, illustrated in c. (d) An overlay of the RFP indicator and BIND scan show that detailed morphology of cells can be captured by the BIND scan and is similar to the morphology seen by fluorescent light microscopy of the RFP indicator. (e) Incubated BIND Scanner allows long-term time-lapse imaging of iNLCs. BD, bipolar disorder; iNLC, induced neuronal-like cell; RFP, red fluorescent protein.
Mentions: Skin samples from a clinically phenotyped cohort of patients with bipolar 1 disorder and a history of lithium treatment (n=12) and matched screened healthy control subjects (n=6) were expanded into fibroblast lines and subsequently transformed using recombinant lentiviruses carrying transgenes for miR-9/9*-124, NEUROD2, ASCL1 and MYT1L.9 This transformation results in iNLCs, directly transdifferentiated from the starting fibroblast population. The full duration of time to generate ‘mature' neuronal-like cells that express synaptic markers and fire action potentials as described by Yoo et al.9 is 4–5 weeks. However, due to reduced viability we observed over this prolonged time period of differentiation, and the goal of a more rapid screening process, we elected to measure cellular activity at an earlier time point in the transdifferentiation process to facilitate generating reproducible measurements over larger numbers of iNLCs. This period, 13–14 days posttransduction, occurs after antibiotic selection for cells expressing the transgenes, but before the large amount of cellular attrition. Figure 2a shows the timeline of the neural induction.

Bottom Line: However, efficiently identifying and characterizing relevant neuronal phenotypes in the absence of well-defined pathophysiology remains a challenge.In this study, we collected fibroblast samples from patients with bipolar 1 disorder, characterized by their lithium response (n=12), and healthy control subjects (n=6).We identified a cellular phenotype in reprogrammed neurons using a label-free imaging assay based on a nanostructured photonic crystal biosensor and found that an optical measure of cell adhesion was associated with clinical response to lithium treatment.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Psychiatry, Center for Experimental Drugs and Diagnostics, Massachusetts General Hospital, Boston, MA, USA [2] Center for Human Genetics Research, Massachusetts General Hospital, Boston, MA, USA [3] Stanley Center for Psychiatric Research, Broad Institute of MIT & Harvard, Cambridge, MA, USA.

ABSTRACT
Development of novel treatments and diagnostic tools for psychiatric illness has been hindered by the absence of cellular models of disease. With the advent of cellular reprogramming, it may be possible to recapitulate the disease biology of psychiatric disorders using patient skin cells transdifferentiated to neurons. However, efficiently identifying and characterizing relevant neuronal phenotypes in the absence of well-defined pathophysiology remains a challenge. In this study, we collected fibroblast samples from patients with bipolar 1 disorder, characterized by their lithium response (n=12), and healthy control subjects (n=6). We identified a cellular phenotype in reprogrammed neurons using a label-free imaging assay based on a nanostructured photonic crystal biosensor and found that an optical measure of cell adhesion was associated with clinical response to lithium treatment. This cellular phenotype may represent a useful biomarker to evaluate drug response and screen for novel therapeutics.

Show MeSH
Related in: MedlinePlus