ACS Synthetic Biology,
Journal Year:
2025,
Volume and Issue:
unknown
Published: March 26, 2025
Escherichia
coli
is
a
widely
studied
model
organism
and
an
integral
component
of
the
human
gut
microbiome,
offering
significant
potential
for
bacteria-based
therapeutic
applications.
Despite
this
promise,
engineering
native
E.
strains
remains
challenging.
In
study,
we
employed
chassis-independent
recombinase-assisted
genome
(CRAGE)
technique
to
genetically
engineer
strain
EcAZ-1
probiotic
Nissle
1917
(EcN).
We
successfully
expressed
suite
heterologous
genes,
including
bioluminescent
lux
operon,
green
fluorescent
protein
(GFP),
oxygen-independent
IFP2.0,
in
both
strains.
Optimization
IFP2.0
fluorescence
was
achieved
under
aerobic
anaerobic
conditions
by
coexpressing
heme
oxygenase
gene
and/or
supplementing
chromophore
biliverdin
or
hemin.
Additionally,
engineered
these
biosynthesize
bioactive
compounds
naringenin
mycosporine-like
amino
acids.
This
work
highlights
as
versatile
platforms
synthetic
biology,
paving
way
innovative
applications
biomedical
research
development.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2025,
Volume and Issue:
unknown
Published: March 1, 2025
Abstract
Background
The
gut
microbiome
is
intimately
connected
to
cardiovascular
health
through
the
gut-heart
axis
and
plays
a
pivotal
role
in
maintaining
homeostasis.
Myocardial
infarction
(MI)
disrupts
this
homeostatic
balance,
leading
widespread
adverse
effects.
Hyperglycemia,
hallmark
of
metabolic
dysfunction,
further
exacerbates
these
disruptions,
emphasizing
need
understand
underlying
mechanisms
develop
effective
therapeutic
strategies
for
mitigating
cascading
complications
along
axis.
This
study
aims
elucidate
dynamics
barrier
disruption
during
MI,
explore
liver’s
function
as
an
immune
sentinel
process,
with
focus
on
impact
hyperglycemia
microbial
dissemination,
systemic
inflammation,
liver
function.
Methods
A
murine
MI
model
was
used
evaluate
permeability,
bacterial
translocation,
hepatic
responses.
induced
via
permanent
left
anterior
descending
artery
ligation.
Hyperglycemia
established
streptozotocin
injections
high-fat,
high-sugar
diet.
Gut
integrity
assessed
using
FITC-dextran
assays,
translocation
tracked
intravital
imaging
anaerobic
cultures
from
multiple
organs.
Hepatic
analyzed
flow
cytometry,
cytokine
profiling,
phagocytosis
assays.
16S
rRNA
sequencing
characterized
composition
translocated
bacteria.
Results
significantly
increased
intestinal
exacerbating
dysfunction.
Intravital
revealed
portal
vein
liver,
highlighting
interception.
impaired
macrophage
by
activating
NLRP3
inflammasome
signaling,
reducing
clearance
promoting
persistent
colonization.
Systemic
inflammatory
cytokines,
particularly
TNF-α,
were
elevated,
facilitating
dissemination.
demonstrated
host-dependent
stochastic
variability
composition.
Conclusion
serves
key
regulator
gut-liver-heart
but
functionally
compromised
under
hyperglycemia,
inflammation
dissemination
post-MI.
Targeting
signaling
restoring
may
mitigate
post-MI
complications,
hyperglycemic
conditions.
These
findings
underscore
integrated
incorporating
control
microbiome-targeted
interventions
improve
outcomes.
ACS Synthetic Biology,
Journal Year:
2025,
Volume and Issue:
unknown
Published: March 26, 2025
Escherichia
coli
is
a
widely
studied
model
organism
and
an
integral
component
of
the
human
gut
microbiome,
offering
significant
potential
for
bacteria-based
therapeutic
applications.
Despite
this
promise,
engineering
native
E.
strains
remains
challenging.
In
study,
we
employed
chassis-independent
recombinase-assisted
genome
(CRAGE)
technique
to
genetically
engineer
strain
EcAZ-1
probiotic
Nissle
1917
(EcN).
We
successfully
expressed
suite
heterologous
genes,
including
bioluminescent
lux
operon,
green
fluorescent
protein
(GFP),
oxygen-independent
IFP2.0,
in
both
strains.
Optimization
IFP2.0
fluorescence
was
achieved
under
aerobic
anaerobic
conditions
by
coexpressing
heme
oxygenase
gene
and/or
supplementing
chromophore
biliverdin
or
hemin.
Additionally,
engineered
these
biosynthesize
bioactive
compounds
naringenin
mycosporine-like
amino
acids.
This
work
highlights
as
versatile
platforms
synthetic
biology,
paving
way
innovative
applications
biomedical
research
development.
