Recombinant bacterium to decrease tumor growth
View Patent ↗A recombinant bacterium capable of reducing tumor growth is provided, wherein said recombinant bacterium is capable of: a. increased expression of a nucleic acid encoding a chemoreceptor that directs chemotaxis towards tumors, b. accumulation in a quiescent tumor, c. hyper-invasion of a tumor, d. reduced fitness in normal tissue, e. enhanced stimulation of the host innate immune responses, f. delivering a tumor specific DNA vaccine vector to a tumor cell, and g. increased bacterium-induced host programmed cell death.
1. A recombinant Salmonella bacterium, wherein the bacterium has:
a. ΔP murA ::TT araC P BAD murA,
b. at least one mutation selected from the group consisting of ΔP murA ::TT araC P BAD murA and ΔasdA::TT araC P BAD c2, and, optionally, at least one mutation selected from the group consisting of Δ(gmd-fcl) and ΔrelA,
c. constitutive expression, from a constitutive promoter, of at least one nucleic acid selected from the group consisting of tar and tsr,
d. a deletion or mutation to decrease expression of trg,
e. constitutive expression, from a constitutive promoter, of hilA,
f. at least one mutation selected from the group consisting of ΔpurA and Δ(wza-wcaM),
g. regulated delayed synthesis of at least one nucleic acid selected from the group consisting of sopE2 and sopB, and
h. increased expression of at least one nucleic acid selected from the group consisting of sseL tlpA, and avrA and/or expression of at least one protein selected from the group consisting of Fas ligand (FasL) and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL).
2. A bacterium of claim 1 , wherein the bacterium comprises the following mutations: ΔasdA::TT araC P BAD c2, ΔP murA ::TT araC P BAD murA, Δ(wza-wcaM), ΔrelA::araC P BAD lacl TT, Δ(araC P BAD )::P22 P R araBAD, ΔpagP::P lpp IpxE, ΔendA.
3. A bacterium of claim 1 , wherein the bacterium comprises the following mutations: ΔasdA::TT araC P BAD c2, ΔP murA ::TT araC P BAD murA, Δ(wza-wcaM), ΔrelA::araC P BAD lacl TT, Δ(araC P BAD )::P22 P R araBAD, ΔpagP::P lpp IpxE, ΔendA, ΔP tar ::P trc ΔlacO tar.
4. A bacterium of claim 1 , wherein the bacterium comprises the following mutations: ΔasdA::TT araC P BAD c2, ΔP murA ::TT araC P BAD murA, Δ(wza-wcaM), ΔrelA::araC P BAD lacl TT, Δ(araC P BAD )::P22 P R araBAD, ΔpagP::P lpp IpxE, ΔendA, ΔP tar ::P trc ΔlacO tar, ΔP tsr ::P trc ΔlacO tsr.
5. A bacterium of claim 1 , wherein the bacterium comprises the following mutations: ΔasdA::TT araC P BAD c2, ΔP murA ::TT araC P BAD murA, Δ(wza-wcaM), ΔrelA::araC P BAD lacl TT, Δ(araC P BAD )::P22 P R araBAD, ΔpagP::P lpp IpxE, ΔendA, ΔP tar ::P trc ΔlacO tar, ΔP tsr ::P trc ΔlacO tsr, Δtrg or ΔP trg ::rhaRS-P rhaB trg.
6. A bacterium of claim 1 , wherein the bacterium comprises the following mutations: ΔasdA::TT araC P BAD c2, ΔP murA ::TT araC P BAD murA, Δ(wza-wcaM), ΔrelA::araC P BAD lacl TT, Δ(araC P BAD )::P22 P R araBAD, ΔpagP::P lpp IpxE, ΔendA, ΔP tar ::P trc ΔlacO tar, ΔP tsr :P trc ΔlacO tsr, Δtrg or ΔP trg ::rhaRS-P rhaB trg, ΔP hilA ::P trc ΔlacO hilA.
7. A bacterium of claim 1 , wherein the bacterium comprises the following mutations: ΔasdA::TT araC P BAD c2, ΔP murA ::TT araC P BAD murA, Δ(wza-wcaM), ΔrelA::araC P BAD lacl TT, Δ(araC P BAD )::P22 P R araBAD, ΔpagP::P lpp IpxE, ΔendA, ΔP tar ::P trc ΔlacO tar, ΔP tsr ::P trc ΔlacO tsr, Δtrg or ΔP trg ::rhaRS-P rhaB trg, ΔP hilA ::P trc ΔlacO hilA, ΔpurA.
8. A bacterium of claim 1 , wherein the bacterium comprises the following mutations: ΔasdA::TT araC P BAD c2, ΔP murA ::TT araC P BAD murA, Δ(wza-wcaM), ΔrelA::araC P BAD lacl TT, Δ(araC P BAD )::P22 P R araBAD, ΔpagP::P lpp IpxE, ΔendA, ΔP tar ::P trc ΔlacO tar, ΔP tsr ::P trc ΔlacO tsr, Δtrg or ΔP trg ::rhaRS-P rhaB trg, ΔP hilA ::P trc ΔlacO hilA, ΔpurA, ΔP sopE2 ::P trc sopE2.
9. A bacterium of claim 1 , wherein the bacterium comprises the following mutations: ΔasdA::TT araC P BAD c2, ΔP murA ::TT araC P BAD murA, Δ(wza-wcaM), ΔrelA::araC P BAD lacl TT, Δ(araC P BAD )::P22 P R araBAD, ΔpagP::P lpp IpxE, ΔendA, ΔP tar ::P trc ΔlacO tar, ΔP tsr ::P trc ΔlacO tsr, Δtrg or ΔP trg ::rhaRS-P rhaB trg, ΔP hilA ::P trc ΔlacO hilA, ΔpurA, ΔP sopE2 ::P trc sopE2, ΔP tlpA ::P ansB tlpA.
10. A method of inhibiting tumor growth, the method comprising administering a recombinant bacterium of claim 1 to a tumor.
11. A method of treating cancer in a subject, the method comprising administering a recombinant bacterium of claim 1 to the subject, wherein the subject has cancer.
12. A recombinant Salmonella bacterium, wherein the bacterium has:
a. constitutive expression, from a constitutive promoter, of Tar and Tsr, and
b. elimination of expression of Trg.
13. A recombinant Salmonella bacterium, wherein the bacterium has:
a. constitutive expression, from a constitutive promoter, of at least one nucleic acid selected from the group consisting tar and tsr,
b. a deletion or mutation to prevent or decrease expression of trg,
c. constitutive expression, from a constitutive promoter, of hilA,
d. at least one mutation selected from the group consisting of ΔpurA and Δ(wza-wcaM), and
e. increased expression of at least one nucleic acid selected from the group consisting of sseL, tlpA, and avrA and/or a tumor specific DNA vaccine vector that expresses at least one protein selected from the group consisting of FasL and TRAIL.
14. The bacterium of claim 13 , wherein the bacteria further has regulated delayed synthesis of at least one nucleic acid selected from the group consisting of sopE2 and sopB.
15. A recombinant Salmonella bacterium, wherein the bacterium comprises:
a. ΔP tar ::P trc ΔlacO tar and ΔP tsr ::P trc ΔlacO tsr,
b. Δtrg or ΔP trg ::rhaRS-P rhaB trg,
c. ΔP hilA ::ΔP trc ΔlacO hilA,
d. Δ(wza-wcaM) and, optionally, ΔpurA, and
e. ΔP tlpA ::P ansB tlpA or a tumor specific DNA vaccine vector to a tumor cell that expresses FasL.
16. The bacterium of claim 15 , wherein the bacteria further comprises ΔP sopE2 ::P trc sopE2.
17. A recombinant Salmonella bacterium, wherein the bacterium comprises:
a. constitutive expression, from a constitutive promoter, of at least one nucleic acid selected from the group consisting tar and tsr,
b. deletion or mutation to decrease expression of trg,
c. constitutive expression, from a constitutive promoter, of hilA,
d. at least one mutation selected from the group consisting of ΔpurA and Δ(wza-wcaM), and
e. increased expression of at least one nucleic acid selected from the group consisting of sseL, tlpA, and avrA and/or a tumor specific DNA vaccine vector that expresses at least one protein selected from the group consisting of FasL and TRAIL.
18. The bacterium of claim 17 , wherein the bacteria further comprises regulated delayed synthesis of at least one nucleic acid selected from the group consisting of sopE2 and sopB.