Granted Patent B2

US 12,748,102 · App. 18/026,907 · Granted Sep 29, 2026

Inventors: Samuel Sances (Santa Monica, CA); Clive Svendsen (Pacific Palisades, CA)

Assignee: Cedars-Sinai Medical Center

G01N33/5058B01L3/502715C12M21/08C12N5/0619G01N33/5023B01L2300/0874C12N2501/01C12N2501/13C12N2503/02C12N2506/45C12N2513/00C12N2533/52C12N2533/54C12N2533/90

Abstract

Claims (47)

1 . A method of identifying an agent of interest, comprising:

contacting a quantity of neurons or a quantity of ventralized spinal motor neuron progenitor cells (spNPCs) with a test agent in a top channel of a multi-channel 3-dimenional (3-D) cell culture chip;

adding brain microvascular cells (BMECs) to a bottom channel of the multi-channel 3-D cell culture chip;

adding culture media to the bottom channel, causing the culture media to continuously perfuse through the top or bottom channel;

measuring biomarkers of amyotrophic lateral sclerosis (ALS) comprising one or more parameters relating to the quantity of neurons or the quantity of ventralized spNPCs, the one or more parameters being expression level of one or more genes or one or more proteins; and

identifying the test agent as the agent of interest if the test agent modulates the one or more parameters by increasing or decreasing the expression level of one or more genes as compared to a reference level for each gene, or by increasing or decreasing the expression level of one or more proteins as compared to a reference level for each protein,

wherein the quantity of neurons or the quantity of spNPCs is differentiated from induced pluripotent stem cells (iPSCs) from an ALS patient.

2 . The method of claim 1 , wherein the neurons are spinal motor neurons.

3 . The method of claim 1 , wherein the ALS patient is a young onset amyotrophic lateral sclerosis (YOALS) patient.

4 . The method of claim 3 , wherein the agent of interest is a drug for treating amyotrophic lateral sclerosis (ALS).

5 . The method of claim 3 , wherein the agent of interest is a drug for treating young onset amyotrophic lateral sclerosis (YOALS).

6 . The method of claim 1 , wherein the agent of interest is a drug that can cross the blood-brain barrier.

7 . The method of claim 1 , wherein

the one or more parameters is expression level of one or more genes or one or more proteins, and

the test agent is identified as the agent of interest if the test agent increases or decreases the expression level of one or more genes as compared to a reference level for each gene, or the test agent increases or decreases the expression level of one or more proteins as compared to a reference level for each protein.

8 . The method of claim 7 , wherein

the one or more genes are selected from the group consisting of TOMM40L, BAX, RACI, BAD, DAXX, PRPH2, GPX1, SOD1, PP3R2, CAT, MAP2K6, TNFRSF1A, NEFM, PRPH, NEFL, NEFH, and combinations thereof, and

the test agent is identified as the agent of interest if the test agent increases or decreases the expression level of the one or more genes as compared to a reference level for each gene.

9 . The method of claim 7 , wherein

the one or more genes are selected from the group consisting of neurofilament light (NEFL), neurofilament medium (NEFM), neurofilament heavy (NEFH), peripherin (PRPH), neuronal microtubule gene beta 3 tubulin (TUBB3), and combinations thereof, and

the test agent is identified as the agent of interest if the test agent increases or decreases the expression level of the one or more genes as compared to a reference level for each gene.

10 . The method of claim 7 , wherein

the one or more genes are neurofilament light (NEFL), neurofilament medium (NEFM), neurofilament heavy (NEFH), and peripherin (PRPH), and

the test agent is identified as the agent of interest if the test agent increases or decreases the expression level of the one or more genes as compared to a reference level for each gene.

11 . The method of claim 7 , wherein

the one or more proteins are selected from the group consisting of CCS, SLS1A2, PPP2CB, BCL2L1, RAC1, PPP3R1, SOD1, TOMM40, GPX1, CAT, MAPK14, NEFH, NEFM, NEFL, PRPH, and combinations thereof, and

the test agent is identified as the agent of interest if the test agent increases or decreases the expression level of one or more proteins as compared to a reference level for each protein.

12 . The method of claim 7 , wherein

the one or more proteins are selected from the group consisting of neurofilament light (NFL), neurofilament medium (NFM), neurofilament heavy (NFH), peripherin (PERI), neuronal microtubule gene beta 3 tubulin (TBB3) protein, and combinations thereof, and

the test agent is identified as the agent of interest if the test agent increases or decreases the expression level of one or more proteins as compared to a reference level for each protein.

13 . The method of claim 7 , wherein

the one or more proteins are neurofilament light (NFL), neurofilament medium (NFM), neurofilament heavy (NFH), and peripherin (PERI), and

the test agent is identified as the agent of interest if the test agent increases or decreases the expression level of one or more proteins as compared to a reference level for each protein.

14 . The method of claim 1 , further comprising treating ALS patients with the identified agent of interest.

15 . The method of claim 14 , wherein the identified agent of interest is capable of crossing the blood-brain barrier.

16 . The method of claim 1 , wherein the quantity of neurons or the quantity of spNPCs differentiated from iPSCs from the ALS patients exhibits a significant increase in neurofilament heavy (NEFH) transcripts and neurofilament heavy (NFH) protein in the 3-D cell culture chip, compared to non-diseased control lines.

17 . The method of claim 1 , wherein the BMECs are added to the bottom channel 14 days after adding the quantity of neurons or the quantity of spNPCs to the top channel.

18 . A system, comprising:

a multi-channel 3-dimenional (3-D) cell culture chip having:

a top channel including a quantity of neurons or a quantity of ventralized spinal motor neuron progenitor cells (spNPCs) for contacting a test agent; and

a bottom channel including brain microvascular cells (BMECs) and cell culture media,

wherein the quantity of neurons or the quantity of spNPCs is differentiated from induced pluripotent stem cells (iPSCs) from an amyotrophic lateral sclerosis (ALS) patient, and

wherein the system is configured to perfuse the cell culture media continuously through the top or bottom channel.

19 . The system of claim 18 , wherein the neurons are spinal motor neurons.

20 . The system of claim 18 , further comprising a test agent.

21 . The system of claim 18 , wherein the ALS patient is a young onset amyotrophic lateral sclerosis (YOALS) patient.

22 . The system of claim 18 , wherein the multi-channel 3-D cell culture chip contains increased neurofilament expression.

Assignments (1)

Continuity (2)

References Cited (400)