Biosensor tattoos and uses therefor for biomarker monitoring

Patent metadata

Field	Value
Publication number	US-12420073-B2
Application number	US-202017132511-A
Country	US
Kind code	B2
Filing date	Dec 23, 2020
Priority date	Apr 23, 2015
Publication date	Sep 23, 2025
Grant date	Sep 23, 2025

Abstract

Provided herein are devices and methods used to produce tattoo biosensors that are based on spatially controlled intracutaneous gene delivery of optical reporters driven by specific transcription factor pathways for a given cytokine or other analyte. The biosensors can be specific to a given analyte, or more generically represent the convergence of several cytokines into commonly shared intracellular transcription factor pathways. These biosensors can be delivered as an array in order to monitor multiple cytokines. Biosensor redeployment can enable chronic monitoring from months to years. The tattooed biosensor array of the present invention includes endogenous reporter cells, naturally tuned to each patient's own biology and can be used to reliably measure the state of a patient in real-time.

First claim

We claim:

1. A microneedle array comprising:
a backing; and
a plurality of microneedles attached to a side of the backing and comprising:
a first nucleic acid comprising a first gene encoding a colorimetric protein under transcriptional control of a vertebrate transcription factor-responsive element (TRE) such that when transfected into a vertebrate cell, the gene is expressed differently in the presence of a vertebrate transcription factor that binds the TRE than in the absence of the transcription factor and the difference in expression of the gene is optically detectable, wherein the first nucleic acid is packaged in an Adeno-associated virus (AAV) transducing particle,
one or more additional, different nucleic acids, with each of the one or more additional, different nucleic acids comprising a second gene encoding a colorimetric protein that is the same or different from the colorimetric protein of the first gene, responsive to the same or different transcriptional control than the first gene, and
a reference reporter gene under transcriptional control of an inducible promotor and encoding a colorimetric protein that is the same or different from the colorimetric protein of the first gene and the second gene,

wherein the microneedles containing the nucleic acids of the first gene, the second gene, and the reference reporter gene are located in discrete, addressable locations within individual or clusters of microneedles, such that when deposited in the skin of a patient, a colorimetric response is spatially patterned.

2. The microneedle array of claim 1 wherein a single microneedle comprises two or more of the different nucleic acids that produce different, separately addressable colorimetric proteins.

3. The microneedle array of claim 2 , wherein the separately addressable colorimetric proteins are two different fluorescent proteins that have distinguishable excitation and/or emission spectra.

4. The microneedle array of claim 1 , wherein at least a portion of the microneedles comprising the nucleic acids, are dissolvable in vivo.

5. The microneedle array of claim 1 , wherein the first nucleic acid is a self-complementary AAV genome.

6. The microneedle array of claim 1 , wherein the colorimetric protein is a fluorescent protein.

7. The microneedle array of claim 6 , wherein which the fluorescent protein is a green, yellow, cyan, red, far-red or near-infrared fluorescent protein.

8. The microneedle array of claim 7 , wherein the fluorescent protein is a far-red or near-infrared fluorescent protein, and the far-red or near-infrared fluorescent protein is one of eqFP578, Katushka, mKate, mNeptune, e2-Crimson, TagRFP657, mCardinal, iRFP670, iRFP682, iRFP702, iRFP713, iRFP720, iSplit, PAiRFP1, PAiRFP2, mCherry, tdTomato, DsRed-Monomer, dsRed-Express2, dsRed-Express, dsRed2, asRed2, mStrawberry, mRuby, mApple, jRed, HcRed1, mRaspberry, dKeima-Tandem, mPlum, AQ143, mIFP, iFP1.4, iFP2.0, or NirFP.

9. The microneedle array of claim 1 , in which the first gene is under transcriptional control of a transcription control sequence comprising the TRE.

10. The microneedle array of claim 9 , wherein the transcription control sequence comprises a minimal cytomegalovirus (CMV) promoter 3′ to the TRE.

11. The microneedle array of claim 1 , wherein the first gene is under transcriptional control of a TRE chosen from: AP-1 TRE, C/EBPalpha TRE, c-Fos TRE, c-Jun TRE, c-Myc TRE, c-Rel TRE, DP-1 TRE, E2F+p107 TRE, E2F-1 TRE, E2F-4/DP-2 TRE, Egr-1 TRE, ErbA TRE, FosB TRE, HIF-1 TRE, HSF1 TRE, INF TRE, JunD TRE, Max1 TRE, NF-κB TRE, N-Myc TRE, p53 TRE, REVERB-alpha TRE, Sp1 TRE, Sp3 TRE, SRF TRE, YY1 TRE, NFAT TRE, FOXO1 TRE, ETS-1 TRE, RELA TRE, STAT1 TRE, STAT2 TRE, STAT1/2 TRE, STAT3 TRE, CREB TRE, IRF1 TRE, and/or SRC-1 TRE.

12. The microneedle array of claim 1 , wherein the second gene is under transcriptional control of a TRE different from that of the first gene, the TRE chosen from one or more of AP-1 TRE, C/EBPalpha TRE, c-Fos TRE, c-Jun TRE, c-Myc TRE, c-Rel TRE, DP-1 TRE, E2F+p107 TRE, E2F-1 TRE, E2F-4/DP-2 TRE, Egr-1 TRE, ErbA TRE, FosB TRE, HIF-1 TRE, HSF1 TRE, INF TRE, JunD TRE, Max1 TRE, NF-κB TRE, N-Myc TRE, p53 TRE, REVERB-alpha TRE, Sp1 TRE, Sp3 TRE, SRF TRE, YY1 TRE, NFAT TRE, FOX01 TRE, ETS-1 TRE, RELA TRE, STAT1 TRE, STAT2 TRE, STAT1/2 TRE, STAT3 TRE, CREB TRE, IRF1 TRE, and/or SRC-1 TRE.

13. The microneedle array of claim 1 , wherein the nucleic acids are placed in layers in the microneedles at a distance from the backing of at least 50 μm.

14. The microneedle array of claim 1 , wherein the nucleic acids are placed in layers in the microneedles, and wherein the microneedles comprise one or more additional nucleic acids, or nucleic acids comprising reporter genes, and optionally constitutive or control reporter genes, drugs, or excipients placed in one or more additional layers.

15. A microneedle array comprising:
a backing;
a plurality of microneedles attached to a side of the backing and extending away from the backing, each of the plurality of microneedles comprising a stem adjacent to the backing and a tip spaced from the backing and comprising at least a first layer arranged a first distance from the backing and a second layer arranged a second distance from the backing;
a first nucleic acid, received within the first layer of the tip of one or more of the plurality of microneedles, the first nucleic acid comprising a first gene encoding a colorimetric protein under transcriptional control of a vertebrate transcription factor-responsive element (TRE) such that when transfected into a vertebrate cell, the gene is expressed differently in the presence of a vertebrate transcription factor that binds the TRE than in the absence of the transcription factor and the difference in expression of the gene is optically detectable, wherein the first nucleic acid is packaged in a viral particle; and
one or more additional, different nucleic acids received within the second layer of the tip of one or more of the plurality of microneedles, each of the one or more additional, different nucleic acids comprising a second gene encoding a colorimetric protein that is the same or different from the colorimetric protein of the first gene, responsive to the same or different transcriptional control than the first gene.

16. The microneedle array of claim 15 , wherein the microneedles containing the nucleic acids of the first gene and the second gene are located in discrete, addressable locations within individual or clusters of microneedles, such that when deposited in the skin of a patient, a colorimetric response is spatially patterned.

17. The microneedle array of claim 15 , wherein the tip comprises a bioerodible material that dissolves in about 30 minutes in vivo.

18. The microneedle array of claim 15 , wherein the stem and the tip of the plurality of microneedles are formed of a bioerodible material, and wherein the bioerodible material of the stem dissolves more quickly than the bioerodible material of the tip.

19. The microneedle array of claim 15 , wherein the first layer and the second layer are formed of different bioerodible materials, such that the first layer dissolves at a different rate than the second layer.

Assignees

• Univ Carnegie Mellon
• Univ Of Pittsburgh—Of The Commonwealth System Of Higher Education
• Univ Of Pittsburgh—Of The Commonwealth Systems Of Higher Education

Inventors

• Ozdoganlar O Burak
• Bruchez Marcel P
• Campbell Phil G
• Jarvik Jonathan W
• FALO Louis
• Erdos Geza

Classifications

• A61M2037/0061

  Methods for using microneedles · CPC title

• A61M37/0084

  Tattooing apparatus with incorporated liquid feeding device · CPC title

• A61K49/0097

  Cells, viruses, ghosts, red blood cells, viral vectors, used for imaging or diagnosis in vivo · CPC title

• A61K49/0045

  the fluorescent agent being a peptide or protein used for imaging or diagnosis in vivo · CPC title

• A61K49/0006

  Skin tests, e.g. intradermal testing, test strips, delayed hypersensitivity · CPC title