COMPOSITIONS AND METHODS FOR TREATING DISEASES AND DISORDERS USING FOURNIERELLA MASSILIENSIS

1 . A pharmaceutical composition comprising Fournierella massiliensis bacteria.

2 . The pharmaceutical composition of claim 1 , wherein the Fournierella massiliensis is a strain comprising at least 90% genomic, 16S, and/or CRISPR sequence identity to the nucleotide sequence of the Fournierella massiliensis Strain A (ATCC Deposit Number PTA-126696).

3 . The pharmaceutical composition of claim 1 , wherein the Fournierella massiliensis is a strain comprising at least 99% genomic, 16S, and/or CRISPR sequence identity to the nucleotide sequence of the Fournierella massiliensis Strain A (ATCC Deposit Number PTA-126696).

4 . The pharmaceutical composition of claim 1 , wherein the Fournierella massiliensis is a strain comprising at least 99% 16S sequence identity to SEQ ID NO: 1.

5 . The pharmaceutical composition of claim 1 , wherein the Fournierella massiliensis is Fournierella massiliensis Strain A (ATCC Deposit Number PTA-126696).

6 . The pharmaceutical composition of any one of claims 1-5 , wherein at least 50% of the bacteria in the pharmaceutical composition are Fournierella massiliensis Strain A.

7 . The pharmaceutical composition of any one of claims 1-6 , wherein at least 90% of the bacteria in the pharmaceutical composition are Fournierella massiliensis Strain A.

8 . The pharmaceutical composition of any one of claims 1-7 , wherein substantially all of the bacteria in the pharmaceutical composition are Fournierella massiliensis Strain A.

9 . The pharmaceutical composition of any one of claims 1-8 , wherein the bacterial composition comprises at least 1 × 10 6 total cells of Fournierella massiliensis Strain A.

10 . The pharmaceutical composition of any one of claims 1-9 , wherein the bacterial composition comprises at least 1 × 10 7 total cells of Fournierella massiliensis Strain A.

11 . The pharmaceutical composition of any one of claims 1-10 , wherein the bacterial composition comprises at least 1 × 10 8 total cells of Fournierella massiliensis Strain A.

12 . The pharmaceutical composition of any one of claims 1-11 , wherein the pharmaceutical composition comprises live bacteria.

13 . The pharmaceutical composition of any one of claims 1-11 , wherein the pharmaceutical composition comprises attenuated bacteria.

14 . The pharmaceutical composition of any one of claims 1-11 , wherein the pharmaceutical composition comprises killed bacteria.

15 . The pharmaceutical composition of any one of claims 1-14 , wherein the pharmaceutical composition comprises lyophilized bacteria.

16 . The pharmaceutical composition of any one of claims 1-15 , wherein the pharmaceutical composition comprises irradiated bacteria.

17 . The pharmaceutical composition of claim 16 , wherein the pharmaceutical composition comprises gamma irradiated bacteria.

18 . A pharmaceutical composition comprising isolated extracellular vesicles (mEVs) produced from Fournierella massiliensis .

19 . The pharmaceutical composition of claim 18 , wherein the Fournierella massiliensis is a strain comprising at least 90% genomic, 16S, and/or CRISPR sequence identity to the nucleotide sequence of the Fournierella massiliensis Strain A (ATCC Deposit Number PTA-126696).

20 . The pharmaceutical composition of claim 18 , wherein the Fournierella massiliensis is a strain comprising at least 99% genomic, 16S, and/or CRISPR sequence identity to the nucleotide sequence of the Fournierella massiliensi s Strain A (ATCC Deposit Number PTA-126696).

21 . The pharmaceutical composition of claim 18 , wherein the Fournierella massiliensis is a strain comprising at least 99% 16S sequence identity to SEQ ID NO: 1.

22 . The pharmaceutical composition of claim 18 , wherein the Fournierella massiliensis is Fournierella massiliensis Strain A (ATCC Deposit Number PTA-126696).

23 . The pharmaceutical composition of claim 18 , wherein at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% of of the pharmaceutical composition is mEVs.

24 . The pharmaceutical composition of any one of claims 18-23 , wherein the composition comprises secreted mEVs (smEVs).

25 . The pharmaceutical composition of any one of claims 18-23 , wherein the composition comprises processed mEVs (pmEVs).

26 . The pharmaceutical composition of any one of claims 18-23 , wherein the mEVs comprise pmEVs and the pmEVs are produced from bacteria that have been gamma irradiated, UV irradiated, heat inactivated, acid treated or oxygen sparged.

27 . The pharmaceutical composition of any one of claims 18-23 , wherein the mEVs comprise pmEVs and the pmEVs are produced from live bacteria.

28 . The pharmaceutical composition of any one of claims 18-27 , wherein the mEVs are lyophilized (e.g., the lyophilized product further comprises a pharmaceutically acceptable excipient).

29 . The pharmaceutical composition of any one of claims 18-28 , wherein the mEVs are gamma irradiated.

30 . The pharmaceutical composition of any one of claims 18-28 , wherein the mEVs are UV irradiated.

31 . The pharmaceutical composition of any one of claims 18-28 , wherein the mEVs are heat inactivated (e.g., at 50° C. for two hours or at 90° C. for two hours).

32 . The pharmaceutical composition of any one of claims 18-28 , wherein the mEVs are acid treated.

33 . The pharmaceutical composition of any one of claims 18-28 , wherein the mEVs are oxygen sparged (e.g., at 0.1 vvm for two hours).

34 . The pharmaceutical composition of any one of claims 18-33 , wherein the dose of mEVs is about 2×10 6 to about 2×10 16 particles (e.g., wherein particle count is determined by NTA (nanoparticle tracking analysis)).

35 . The pharmaceutical composition of any one of claims 18-34 , wherein the dose of mEVs is about 5 mg to about 900 mg total protein (e.g., wherein total protein is determined by Bradford assay or BCA assay).

36 . A pharmaceutical composition comprising Fournierella massiliensis microbial extracellular vesicles (mEVs) and Fournierella massiliensis bacteria.

37 . The pharmaceutical composition of claim 36 , wherein the Fournierella massiliensis is a strain comprising at least 90% genomic, 16S, and/or CRISPR sequence identity to the nucleotide sequence of the Fournierella massiliensis Strain A (ATCC Deposit Number PTA-126696).

38 . The pharmaceutical composition of claim 36 , wherein the Fournierella massiliensis is a strain comprising at least 99% genomic, 16S, and/or CRISPR sequence identity to the nucleotide sequence of the Fournierella massiliensis Strain A (ATCC Deposit Number PTA-126696).

39 . The pharmaceutical composition of claim 36 , wherein the Fournierella massiliensis is a strain comprising at least 99% 16S sequence identity to SEQ ID NO: 1.

40 . The pharmaceutical composition of claim 36 , wherein the Fournierella massiliensis is Fournierella massiliensis Strain A (ATCC Deposit Number PTA-126696).

41 . The pharmaceutical composition of any one of claims 36-40 , wherein at least, about, or no more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the total particles in the pharmaceutical composition are Fournierella massiliensis mEVs.

42 . The pharmaceutical composition of any one of claims 36-40 , wherein at least, about, or no more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the total particles in the pharmaceutical composition are Fournierella massiliensis bacteria particles.

43 . The pharmaceutical composition of any one of claims 36-40 , wherein at least, about, or no more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the total proteins in the pharmaceutical composition are Fournierella massiliensis mEVs.

44 . The pharmaceutical composition of any one of claims 36-40 , wherein at least, about, or no more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the total proteins in the pharmaceutical composition are Fournierella massiliensis bacteria proteins.

45 . The pharmaceutical composition of any one of claims 36-40 , wherein at least, about, or no more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the total lipids in the pharmaceutical composition are Fournierella massiliensis mEVs.

46 . The pharmaceutical composition of any one of claims 36-40 , wherein at least, about, or no more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the total lipids in the pharmaceutical composition are Fournierella massiliensis bacteria lipids.

47 . The pharmaceutical composition of any one of claims 1-46 , wherein the pharmaceutical composition is for the treatment of a disease.

48 . The pharmaceutical composition of any one of claims 1-47 , wherein the pharmaceutical composition is for the treatment of an immune disorder (e.g., a cancer, an autoimmune disease, an inflammatory disease, a dysbiosis, and/or a metabolic disease).

49 . The pharmaceutical composition of any one of claims 1-47 , wherein the pharmaceutical composition is for the treatment of an inflammatory disorder (e.g., dermatitis).

50 . The pharmaceutical composition of any one of claims 1-47 , wherein the pharmaceutical composition is for the treatment of a cancer (e.g., colorectal cancer).

51 . The pharmaceutical composition of any one of claims 1-47 , wherein the pharmaceutical composition is for the treatment of a dysbiosis.

52 . The pharmaceutical composition of any one of claims 1-51 , wherein the pharmaceutical composition induces an immune response.

53 . The pharmaceutical composition of any one of claims 1-52 , wherein the pharmaceutical composition activates innate antigen presenting cells.

54 . The pharmaceutical composition of any one of claims 1-53 , wherein the pharmaceutical composition is formulated for oral, rectal, sublingual, intradermal, intraperitoneal,or subcutaneous administration.

55 . The pharmaceutical composition of any one of claims 1-54 , wherein the pharmaceutical composition has one or more beneficial immune effects outside the gastrointestinal tract, e.g., when orally administered.

56 . The pharmaceutical composition of any one of claims 1-55 , wherein the pharmaceutical composition modulates immune effects outside the gastrointestinal tract in the subject, e.g., when orally administered.

57 . The pharmaceutical composition of any one of claims 1-56 , wherein the pharmaceutical composition comprises a solid dose form.

58 . The pharmaceutical composition of claim 57 , wherein the solid dose form comprises a tablet, a minitablet, a capsule, a pill, or a powder, or a combination of the foregoing.

59 . The pharmaceutical composition of claim 57 or 58 , wherein the solid dose form further comprises a pharmaceutically acceptable excipient.

60 . The pharmaceutical composition of any one of claims 57-59 , wherein the solid dose form comprises an enteric coating.

61 . The pharmaceutical composition of any one of claims 57-60 , wherein the solid dose form is for oral administration.

62 . The pharmaceutical composition of any one of claims 1-56 , wherein the pharmaceutical composition comprises a suspension.

63 . The pharmaceutical composition of claim 62 , wherein the suspension is for oral administration (e.g., the suspension comprises PBS, and optionally, sucrose or glucose).

64 . The pharmaceutical composition of claim 62 , wherein the suspension is for intravenous administration (e.g., the suspension comprises PBS).

65 . The pharmaceutical composition of claim 62 , wherein the suspension is for intraperitoneal administration (e.g., the suspension comprises PBS).

66 . The pharmaceutical composition of claim 62 , wherein the suspension is for intratumoral administration (e.g., the suspension comprises PBS).

67 . The pharmaceutical composition of any one of claims 62-66 , wherein the suspension further comprises a pharmaceutically acceptable excipient.

68 . The pharmaceutical composition of any one of claims 62-67 , wherein the suspension further comprises a buffer (e.g., PBS).

69 . The pharmaceutical composition of any one of claims 1-68 , wherein the composition further comprises one or more additional therapeutic agents.

70 . The pharmaceutical composition of any one of claims 1-69 , wherein the pharmaceutical composition is formulated for a daily dose.

71 . The pharmaceutical composition of any one of claims 1-69 , wherein the pharmaceutical composition is formulated for twice a day dose, wherein each dose is half of the daily dose.

72 . The pharmaceutical composition of any one of claims 1-71 for use in treating a disease (e.g., a cancer, an autoimmune disease, an inflammatory disease, a dysbiosis, and/or a metabolic disease).

73 . Use of a pharmaceutical composition of any one of claims 1-71 for the preparation of a medicament for the treatment of a disease (e.g., a cancer, an autoimmune disease, an inflammatory disease, a dysbiosis, and/or a metabolic disease).

74 . A method of treating a subject (e.g., human) in need thereof, comprising administering to the subject a pharmaceutical composition of any one of claims 1-71 .

75 . The method of claim 74 , wherein the subject is in need of treatment for an immune disorder or a metabolic disorder.

76 . The method of claim 74 , wherein the subject is in need of treatment for a cancer.

77 . The method of claim 74 , wherein the subject is in need of treatment for an inflammatory disease.

78 . The method of claim 74 , wherein the subject is in need of treatment for a dysbiosis.

79 . The method of any one of claims 74-78 , further comprising administering to the subject an additional therapeutic agent.

80 . The method of any one of claims 74-79 , wherein the pharmaceutical composition is administered intravenously.

81 . The method of any one of claims 74-79 , wherein the pharmaceutical composition is administered intratumorally.

82 . The method of any one of claims 74-79 , wherein the pharmaceutical composition is administered subtumorally.

83 . The method of any one of claims 74-79 , wherein the pharmaceutical composition is administered by injection, e.g., subcutaneous, intradermal, or intraperitoneal injection.

84 . The method of any one of claims 74-79 , wherein the pharmaceutical composition is administered orally.

85 . The method of any one of claims 74-84 , wherein the pharmaceutical composition further comprises one or more additional therapeutic agents.

86 . The method of any one of claims 74-85 , wherein the dose of mEVs in the pharmaceutical composition is about 2×10 6 to about 2×10 16 particles (e.g., wherein particle count is determined by NTA (nanoparticle tracking analysis)).

87 . The method of any one of claims 74-86 , wherein the dose of mEVs in the pharmaceutical composition is 5 mg to about 900 mg total protein (e.g., wherein total protein is determined by Bradford assay or BCA assay).

88 . The method of any one of claims 74-87 , wherein the pharmaceutical composition is administered once a day.

89 . The method of any one of claims 74-87 , wherein the pharmaceutical composition is administered twice a day.

90 . The method of any one of claims 74-87 , wherein the pharmaceutical composition is formulated for a daily dose.

91 . The method of any one of claims 74-87 , wherein the pharmaceutical composition is formulated for twice a day dose, wherein each dose is half of the daily dose.

92 . A method for preparing a pharmaceutical composition of any one of claims 1-71 in a suspension, the method comprising: combining mEVs, bacteria, or any combination thereof, with a pharmaceutically acceptable buffer (e.g., PBS); thereby preparing the pharmaceutical composition.

93 . The method of claim 92 , wherein the suspension further comprises sucrose or glucose.

94 . The method of claim 92 or 93 , wherein the suspension is for oral administration.

95 . The method of claim 92 or 93 , wherein the suspension is for intravenous administration.

96 . The method of claim 92 or 93 , wherein the suspension is for intraperitoneal administration.

97 . The method of claim 92 or 93 , wherein the suspension is for intratumoral administration.

98 . The method of any one of claims 92-97 , wherein the suspension further comprises a pharmaceutically acceptable excipient.

99 . The method of any one of claims 92-98 , wherein the suspension further comprises a buffer (e.g., PBS).

100 . The method of any one of claims 92-99 , wherein the composition further comprises one or more additional therapeutic agents.

101 . The method of any one of claims 92-100 , wherein the pharmaceutical composition is administered orally.

102 . The method of any one of claims 92-100 , wherein the pharmaceutical composition is administered intravenously.

103 . The method of any one of claims 92-100 , wherein the pharmaceutical composition is administered intratumorally.

104 . The method of any one of claims 92-100 , wherein the pharmaceutical composition is administered subtumorally.

105 . The method of any one of claims 92-100 , wherein the pharmaceutical composition is administered by injection, e.g., subcutaneous, intradermal, or intraperitoneal injection.

106 . The method of any one of claims 92-105 , wherein the dose of mEVs in the pharmaceutical composition is about 2×10 6 to about 2×10 16 particles (e.g., wherein particle count is determined by NTA (nanoparticle tracking analysis)).

107 . The method of any one of claims 92-106 , wherein the dose of mEVs in the pharmaceutical composition is 5 mg to about 900 mg total protein (e.g., wherein total protein is determined by Bradford assay or BCA assay).

108 . A pharmaceutical composition prepared by the method of any one of claims 92-107 .

109 . A method for preparing a pharmaceutical composition of any one of claims 1-71 in a solid dose form, the method comprising:

a) combining the mEVs, bacteria, or any combination thereof, of any one of claims 1-71 with a pharmaceutically acceptable excipient, and

b) compressing the mEVs, bacteria, or any combination thereof; and a pharmaceutically acceptable excipient, thereby preparing the pharmaceutical composition.

110 . The method of claim 109 , wherein the method further comprises enterically coating the solid dose form.

111 . The method of claim 109 or 110 , wherein the solid dose form comprises a tablet, a minitablet, a capsule, a pill, or a powder, or a combination of the foregoing.

112 . The method of any one of claims 109-111 , wherein the composition further comprises one or more additional therapeutic agents.

113 . The method of any one of claims 109-112 , wherein the pharmaceutical composition is administered orally.

114 . The method of any one of claims 109-113 , wherein the dose of mEVs in the pharmaceutical composition is about 2×10 6 to about 2×10 16 particles (e.g., wherein particle count is determined by NTA (nanoparticle tracking analysis)).

115 . The method of any one of claims 109-114 , wherein the dose of mEVs in the pharmaceutical composition is 5 mg to about 900 mg total protein (e.g., wherein total protein is determined by Bradford assay or BCA assay).

116 . A pharmaceutical composition prepared by the method of any one of claims 109-115 .