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Engineered commensal bacteria and methods of use

A technology of bacteria and bacterial cells, applied in biochemical equipment and methods, bacteria, antibacterial drugs, etc., can solve problems that have not shown the mechanistic relationship of microbiota

Pending Publication Date: 2020-01-17
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although linking physiological processes to the microbiome is conceptually exciting, the field is still nascent; studies have shown strong associations, but few clear links between the microbiota and various physiological processes have yet been shown. institutional relationship

Method used

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  • Engineered commensal bacteria and methods of use
  • Engineered commensal bacteria and methods of use
  • Engineered commensal bacteria and methods of use

Examples

Experimental program
Comparison scheme
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preparation example Construction

[0058] 1. Preparation method of engineered native / symbiotic bacteria

[0059] The preparation of therapeutically suitable engineered native or commensal bacteria using heterologous polynucleotides for transformation is straightforward.

[0060] Microbiome samples were obtained from patients. A microbiome sample can be from any population of bacteria that stably colonizes a surface or cavity of an individual. Microflora with stable colonized bacterial flora can be found on all environmentally exposed sites of the body, including the skin, nasopharynx, oral cavity, respiratory tract, gastrointestinal tract and / or female genital tract. Thus, in certain embodiments, microorganisms are obtained by swabbing, rinsing, or biopsy of the skin, nasopharyngeal or oral cavity (e.g., cheek, tongue, gums, or larynx), respiratory tract, gastrointestinal tract, and / or genitourinary tract Group samples. In certain embodiments, the microbiome sample is obtained from a stool sample. In certai...

Embodiment 1

[0142] Example 1. Exemplary Methods for the Preparation of Engineered Symbiotic Bacteria

[0143] Sample collection. For human patients, mucosal biopsies of the duodenum, ileum, and colon were taken during endoscopy and suspended in 1.8 mL of 1×PBS in a 15 mL conical tube at room temperature. Within 8 hours, transfer samples to sterile 2 mL screw top microcentrifuge tubes and add sterile 1.5 mm diameter chrome-plated steel beads.

[0144] For mice, collect feces by placing mice in sterile 1 L polypropylene cups until they excrete 2 to 5 whole pellets. Return the mice to their cages and collect the pellet with sterile forceps and place it in a sterile pre-weighed 2 mL screw top microcentrifuge tube. Collect tissue samples after euthanasia by carbon dioxide and cervical dislocation and place them in pre-weighed sterile 2 mL screw-top microcentrifuge tubes. Subsequent processing is the same for tissue and stool. Within 2 h, the tubes were reweighed and 1 mL of sterile deioniz...

Embodiment 2

[0164] Example 2. Colonization of the Mouse Gut by Reintroduced and Engineered Commensal Bacteria

[0165] In this example, an E. coli strain was isolated from C57Bl / 6 mice, engineered to express GFP and the bile salt hydrolase gene from Lactobacillus, and then reintroduced into C57Bl / 6 mice.

[0166] foreword

[0167] Malnutrition is associated with changes in social, communication, stress-related and cognitive behaviors in murine models (11,12). Human studies have linked shifts in the gut microbiome to autism spectrum disorder (13), major depression (14) and Parkinson's disease (12). Accumulating evidence suggests that microbiome-neuroimmune interactions can mediate the behavioral and physiological abnormalities observed in murine models, notably through global changes in the brain transcriptome, changes in microglial maturation and function, and Integrity of the blood-brain barrier (BBB) ​​(1,15). However, it is unclear which agents and by what mechanism these effects ar...

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Abstract

The present invention provides compositions comprising populations of commensal bacteria isolated from a microbiome sample of a mammalian subject and engineered to express a heterologous polynucleotide, compositions comprising such engineered commensal bacteria and methods of use for delivering a therapeutic polypeptide to a mammal, e.g, by administering the engineered commensal / native bacteria.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application 62 / 486,068, filed April 17, 2017, which is expressly incorporated herein by reference in its entirety for all purposes. [0003] Statement of Government Support [0004] This work was supported in part by grant numbers R03DK114536 and 1K08DK10290201A1 from the National Institutes of Health. The government has certain rights in this invention. Background technique [0005] Recent advances in sequencing, mass spectrometry, and bioinformatics have advanced our understanding of the role of the microbiome in host physiological processes including, but not limited to, metabolic, inflammatory, behavioral, and neurological disorders (Vuong et al., Annu Rev Neurosci. (2017) 40:21-49) [also cited in 25271724]. Although linking physiological processes to the microbiome is conceptually exciting, the field is still nascent; studies have sho...

Claims

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Application Information

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IPC IPC(8): A61K35/742A61K38/50A61K9/48A61P3/04A61P3/10A61P35/00A61P1/00A61P1/04A61P31/18A61P31/04A61P1/14A61P13/00A61P1/12A61P25/00A61P3/06A61P15/08A61P13/12
CPCA61K9/4866A61K35/742A61K38/50C12N1/20C12Y305/01024C12N9/80A61P1/00A61P1/04A61P1/12A61P1/14A61P13/00A61P13/12A61P15/08A61P25/00A61P3/04A61P31/04A61P31/18A61P35/00A61P3/06A61P3/10Y02A50/30
Inventor S·布朗A·扎林帕尔J·斯奈德
Owner RGT UNIV OF CALIFORNIA
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