GENETICALLY PROGRAMMABLE PATHOGEN SENSE AND DESTROY

• US 20120027786A1
• Filed: 02/23/2011
• Published: 02/02/2012
• Est. Priority Date: 02/23/2010
• Status: Abandoned Application

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Gupta, Saurabh, Weiss, Ron
• Application Number:
  US13/033,233
• Publication Number:
  US 20120027786A1
• Time in Patent Office: Days
• US Class Current: 424/184.1
• CPC Class Codes: A61K 2035/11 Medicinal preparations comprising living procariotic cells
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/14 Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/21 from Pseudomonadaceae (F)
• CPC Class Codes: C07K 14/245 Escherichia (G)
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli

Abstract

Aspects of the invention relate to compositions and methods for using recombinant cells to sense and destroy specific pathogens.

Claims

1. A cell that recombinantly expresses (1) a detection circuit, (2) optionally a signal amplifying circuit, and (3) a secretion circuit that secretes a factor that specifically recognizes and destroys a specific pathogen.

2. The cell of claim 1, wherein the cell is a bacterial cell, optionally wherein the bacterial cell is an Escherichia coli (E. coli) cell, such as an E. coli Nissle 1917 (EcN) cell.

6. The cell of claim 1, wherein the cell is an algal cell, a fungal cell (including a yeast cell), an insect cell or an animal cell, optionally wherein the cell is a mammalian cell and optionally wherein the cell is a T cell or a B cell.

11. The cell of claim 1, wherein the cell detects one or more molecules produced by one or more specific pathogens, optionally wherein one or more of the specific pathogens is a bacterial pathogen and/or wherein one or more of the molecules produced by one or more of the specific pathogens is a quorum sensing molecule.

14. The cell of claim 11, wherein the bacterial pathogen is a Pseudomonas bacterial pathogen, optionally a Pseudomonas aeruginosa bacterial pathogen.

16. The cell of claim 14, wherein the cell detects 3-oxo-C12-homoserine lactone (3OC12HSL) that is secreted by the Pseudomonas aeruginosa bacterial pathogen, and optionally wherein the cell comprises a transcriptional regulator that regulates expression of one or more genes in response to 3OC12HSL.

24. The cell of claim 11, wherein the bacterial pathogen is a Vibrio bacterial pathogen, optionally a Vibrio cholerae bacterial pathogen.

26. The cell of claim 24, wherein the cell detects CAI-1 that is secreted by the Vibrio cholerae bacterial pathogen, and optionally wherein the cell comprises a signal amplifying circuit that regulates production of a factor that is secreted by the cell to specifically destroy the Vibrio bacterial pathogen.

28. The cell of claim 26, wherein the cell expresses CqsS, LuxU and/or LuxO, optionally wherein CqsS, LuxU and/or LuxO are codon-optimized.

31. The cell of claim 1, wherein the pathogen is a viral pathogen or a fungal pathogen.

37. The cell of claim 1, wherein the factor that is secreted by the cell is a bacteriocin, optionally wherein the bacteriocin is a lysin, and optionally wherein a flagellar gene, or a portion thereof, is expressed as a fusion with the lysin in the cell.

45. The cell of claim 37, wherein the lysin is a colicin or a pyocin, optionally wherein the colicin or pyocin comprises a chimeric recognition domain that is modified such that the colicin or pyocin specifically recognizes and destroys a specific pathogen.

52. The cell of claim 37, wherein the cell secretes a bacteriocin that is specific for V. cholera, optionally wherein the bacteriocin is selected from the group consisting of Morricin 269, Kurstacin 287, Kenyacin 404, Entomocin 420 and Tolworthcin 524.

56. The cell of claim 11, wherein the bacterial pathogen is a Shigella bacterial pathogen or a Salmonella bacterial pathogen, optionally wherein the bacterial pathogen is a Shigella dysenteriae bacterial pathogen, and optionally wherein the cell detects AI-3 and its lambdoid phage.

59. The cell of claim 56, wherein the cell expresses QseC and QseB, one or more molecular mimics of a Shiga toxin receptor, a chimeric LPS in the outer membrane of the cell, the Shigella lambdoid phage specific receptor (YaeT), one or more Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) sequences specific to incoming phage, and/or a Shigella specific bacteriocin.

69. The cell of claim 1, wherein the cell is a component of a probiotic, a pharmaceutical preparation, a vaccine, a food or nutraceutical, a biospray and/or a water supply system.

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75. A method for destroying a specific pathogen, the method comprising,

providing a cell that recombinantly expresses (1) a detection circuit, (2) optionally a signal amplifying circuit, and (3) a secretion circuit that secretes a factor that specifically recognizes and destroys the specific pathogen.

76. The method of claim 75, wherein the cell is a bacterial cell, optionally wherein the bacterial cell is an Escherichia coli (E. coli) cell such as an E. coli Nissle 1917 (EcN) cell.

80. The method of claim 75, wherein the cell is an algal cell, a fungal cell (including a yeast cell), an insect cell or an animal cell, optionally wherein the cell is a mammalian cell and optionally wherein the cell is a T cell or a B cell.

85. The method of claim 75, wherein the cell detects one or more molecules produced by one or more specific pathogens, optionally wherein one or more of the specific pathogens is a bacterial pathogen and/or wherein one or more of the molecules produced by one or more of the specific pathogens is a quorum sensing molecule.

88. The method of claim 85, wherein the bacterial pathogen is a Pseudomonas bacterial pathogen, a Vibrio cholerae bacterial pathogen, a Shigella bacterial pathogen or a Salmonella bacterial pathogen, optionally wherein the bacterial pathogen is a Shigella dysenteriae bacterial pathogen.

143. The method of claim 75, wherein the cell is a component of a probiotic, a pharmaceutical preparation, a vaccine, a food or nutraceutical, a biospray and/or a water supply system.

149. The method of claim 75, wherein the method is a method of treating or preventing a disease, optionally wherein the disease is an infectious disease, and optionally wherein the cell is provided in a therapeutically effective amount to a subject in need thereof.

152. The method of claim 75, wherein the pathogen is within the gastrointestinal tract and/or within the lung of a subject.

154. The method of claim 75, wherein the method is a method of sterilization, optionally wherein the sterilization comprises sterilization of medical equipment.

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156. A method for protecting against a specific pathogen, the method comprising,

administering to a subject in need thereof an effective amount of a composition comprising a cell that recombinantly expresses (1) a detection circuit, (2) optionally a signal amplifying circuit, and (3) a secretion circuit that secretes a factor that specifically recognizes and destroys the specific pathogen.

224. The method of claim 156, wherein the cell is a component of a probiotic, a pharmaceutical preparation, a vaccine, a food or nutraceutical, a biospray and/or a water supply system.

230. The method of claim 156, wherein the method is a method of treating or preventing a disease, optionally wherein the disease is an infectious disease.

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