Transcript 16.40.perra-congress.. - the Future Health Summit

Future Health Summit
LOW DOSE CYTOKINES IN SUPPORTIVE THERAPY FOR CANCER
From research: Up to date pre-clinical evidence
*****
DUBLIN
May 26th, 2016
Alessandro Perra – Scientific Director of GUNA S.p.a.
Low Dose Medicine into the Universities
MASTER BIENNALE DI II LIVELLO
LOW DOSE MEDICINE IN
PSICO-NERUO-ENDOCRINOIMMUNOLOGIA
Applicazioni in Medicina Generale,
nelle Disfunzioni
Neuro-Muscolo-Scheletriche e
nell’ Healthy Aging
© Dipartimento Scientifico Guna S.p.a.
LOW DOSE MEDICINE
Low Dose Medicine integrates state of the art scientific
advances in:
P.N.E.I.
Molecular
Biology
NanoPharmacology
linked to
Quantum
Physics
© Dipartimento Scientifico Guna S.p.a.
LOW DOSE MEDICINE
A NOVEL, INTEGRATED, SYSTEMIC APPROACH TO DISEASES
LOW DOSE
CYTOKINES
LOW DOSE
GROWTH
FACTORS
BIOREGULATING
PHYSIOLOGY
LOW DOSE
NEURO
PEPTIDES
LOW DOSE
HORMONES
© Dipartimento Scientifico Guna S.p.a.
P
N
E
I
HOMEOSTATIC Control Systems and
bi-directional cross-talk
CENTRAL NERVOUS SYSTEM
&
AUTONOMIC SYSTEM
ENDOCRINE SYSTEM
IMMUNE SYSTEM
Ader, R., Psychoneuroimmunology, IV edition, vol. 1 e 2, Academic Press, Amsterdam 2007.
It is the classical text on the matter, pubblished for the first time in 1981.
© Dipartimento Scientifico Guna S.p.a.
STRESS AXIS
Ts
Tc
CD8+
X
Not correct
stimulation of
Reduction of
IFN-γ
Th2
Th1
DOWN-REGULATION OF Th1
•Cohen M, et al. Increased emotional distress in daughters of breast cancer patients is associated with decreased natural cytotoxic activity, elevated
levels of stress hormones and decreased secretion of Th1 cytokines. Int J Cancer. 2002;100(3):347-54.
•Montoro J, et al. Stress and allergy. J Investig Allergol Clin Immunol. 2009;19 Suppl 1:40-7.
•He Y, et al. Psychological stress exerts effects on pathogenesis of hepatitis B via type-1/type-2 cytokines shift toward type-2 cytokine response. PLoS
One. 2014;9(8):e105530.
•Radice E, et al. Low-doses of sequential-kinetic-activated interferon-gamma enhance the ex vivo cytotoxicity of peripheral blood natural killer cells
from patients with early-stage colorectal cancer. A preliminary study Intern. Immunopharm 2014;19:66-73.
© Dipartimento Scientifico Guna S.p.a.
P.N.E.I. NETWORK AND SIGNALING MOLECULES
AN ALTERATED CROSS-TALK DUE TO AN
UNBALANCE* BETWEEN SIGNALING MOLECULES
PLAYS A CRUCIAL ROLE IN THE ONSET OF
INFLAMMATORY, ALLERGIC AND AUTO-IMMUNE
DISEASES
*ALTERATED CONCENTRATIONS
© Dipartimento Scientifico Guna S.p.a.
DISEASE
10-6
HYPER-CONCENTRATION
HEALTH
PHYSIOLOGICAL CONCENTRATION
10-15
C O P E
Cytokines & Cells Online Pathfinder Encyclopedia
Version 26.7 (Spring 2011 Edition)
HYPO-CONCENTRATION
DISEASE
© Dipartimento Scientifico Guna S.p.a.
g (gram)= 1
DEFINITIONS
10 -1 = 0.1
10 -2 = 0.01
mg (milligram)= 10 -3 = 0.001
μg (microgram)= 10 -6 = 0.000001
ng (nanogram)= 10 -9 = 0.000000001
pcg (picogram)= 10 -12 = 0.000000000001
fg (fentogram)= 10 -15 = 0.000000000000001
© Dipartimento Scientifico Guna S.p.a.
UP- AND DOWN-REGULATION
The membrane receptor plays a KEY role.
ONLY physiological concentrations are able to
activate or reactivate the membrane receptors and
consequently, stimulate the physiological function
of a target cell.
- Akahoshi T et al. Interleukin 1 stimulates its own receptor expression on human fibroblasts through the endogenous
production of prostaglandin(s). J Clin Invest. 1988 Oct;82(4):1219-24
- Samanta AK et al. Interleukin 8 (monocyte-derived neutrophil chemotactic factor) dynamically regulates its own receptor
expression on human neutrophils. J Biol Chem. 1990 Jan 5;265(1):183-9.
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P
N
E
I
HOMEOSTATIC Control Systems and
bi-directional cross-talk
CENTRAL NERVOUS SYSTEM
&
AUTONOMIC SYSTEM
ENDOCRINE SYSTEM
IMMUNE SYSTEM
Ader, R., Psychoneuroimmunology, IV edition, vol. 1 e 2, Academic Press, Amsterdam 2007.
It is the classical text on the matter, pubblished for the first time in 1981.
© Dipartimento Scientifico Guna S.p.a.
Th-1/Th-2 BALANCE
Th1 UP-REGULATION
Th2 UP-REGULATION
Th1
Th17
Crohn’s Disease
Psoriasis
Vitiligo
…
Th2
Th2
Th2 DOWN-REGULATION
Asthma
Atopy
…
Th1
TReg
Th1 DOWN-REGULATION
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Cytokines UP
• IL-1
• IL-6
• TNF-α
• IL-17
• INF-γ
• IL-2
Physiological
• IL-8
concentration
• IL-1
• IL-6
• TNF-α
• IL-17
• INF-γ
• IL-2
• IL-8
Cytokines DOWN
• IL-1
• IL-6
• TNF-α
• IL-17
• INF-γ
• IL-2
• IL-8
HYPER
HEALTH
HYPO
Cytokines DOWN
• TGF-β
• IL-4
• IL-10
Physiological concentration
• TGF-β
• IL-4
• IL-10
Cytokines UP
• TGF-β
• IL-4
• IL-10
© Dipartimento Scientifico Guna S.p.a.
Neither good nor bad in Nature
Raphael I et al. T cell subsets and their signature cytokines in autoimmune and inflammatory diseases. Cytokine (2014),
http://dx.doi.org/10.1016/j.cyto.2014.09.011
IF DISEASES ARE EXPRESSIONS, CONSEQUENCES OF
CHANGED CONCENTRATION OF MESSENGER MOLECULES…
PROBLEM
Is it possible to modulate the
action of cytokines and other
signaling molecules?
© Dipartimento Scientifico Guna S.p.a.
The concept of BALANCE and the use of SKA
low dose cytokines
Antagonistic cytokines are utilized in order
to slow down a biological effect; Same
cytokines in order to enhance a biological
function.
© Dipartimento Scientifico Guna S.p.a.
THE CONCEPT OF BALANCE – RECIPROCITY of TH CELLS
Th0
IL-4 UP-REGULATES
IL-12 UP-REGULATES
Th1
Inflammatory diseases
IL-12, INF-γ
Th2
Allergy
IL-4
IL-4 DOWN-REGULATES
INF-γ DOWN-REGULATES
Th subsets cross-regulate expansion and functions each other.
- Cooke , A. Th17 in Inflammatory Conditions. 2006, Rev Diabetic Stud 3: 72-7
- Bettelli E. et al. Th17: the third member of the effector T cell trilogy. Current Opinion in Immunology 2007, 19: 652-657
© Dipartimento Scientifico Guna S.p.a.
© Dipartimento Scientifico Guna S.p.a.
IL-10 4C
A novel, systemic,
integrated,
synergistic approach for
supportive therapy in
Cancer
© Dipartimento Scientifico Guna S.p.a.
D’Amico L, Ruffini E, Ferracini R, Roato I (2012) Low Dose
of IL-12 stimulates T Cell response in cultures of PBMCs
derived from Non Small Cell Lung Cancer Patients.
Journal of Cancer Therapy 3: 337-342.
© Dipartimento Scientifico Guna S.p.a.
Plasticity and Rigidity of
Relationship among Th1-Th2-Th3
Th0
Th1
Th2
DOWN-REGULATES
DOWN-REGULATES
TGF-β
TReg
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5
4.5
4.5
4
CD4/Treg
3.5
3
day 5
day 10
2.5
2
1.7
1.8
1.7
1.5
0.9
1
0.8
0.6
0.4
0.5
0.2
0
healthy
ctrl
basale
1pg/mlCH3
0.01pg/ml
CH4 SKA
basal
placebo
1010ng/ml
ng/ml
IL12
© Dipartimento Scientifico Guna S.p.a.
%CD4/IFN gamma
Day 5
60
50
40
30
20
10
0
Series1
basale
30.7
CH3
19
CH4
48.4
placebo
48.3
IL12 10 ng/ml
4.8
%CD4/IFN gamma
Day 10
60
50
40
30
20
10
0
Series1
CH3 SKA
IL-12 1pg/ml
32.8
CH4 SKA
IL-12 0.01pg/ml
56
placebo
46.2
IL12 10 ng/ml
4.2
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© Dipartimento Scientifico Guna S.p.a.
Legenda:
Healthy
subjects
Nonmetastatic
cancer
patients
Metastatic
cancer
Non metastatic
Healthy subjects patients
cancer patients
•Left column =
not-treated control group
•Central column =
patients treated with
conventional dosage of
IFN-gamma (1 ng/ml)
•Right column = patients
treated with IFN-gamma
Metastatic
cancer
GUNA (0.25
fg/ml)
patients
© Dipartimento Scientifico Guna S.p.a.
Radice E, Bellone G, Miranda V.
Enhancement of the immunomodulatory functions of ex vivo-generated dendritic cells from earlystage colon cancer patients by consecutive exposure to low doses of sequential kinetic activated IL-4
and IL-12.
A preliminary study.
Translational Oncology 2015;8:327-38
© Dipartimento Scientifico Guna S.p.a.
STUDY TYPE
Ex vivo study on dendritic cells obtained from the peripheral
blood of patients with early stage colorectal carcinoma (CRC) - in
the presence or absence of metastasis - and from healthy donors.
AIM OF THE STUDY
The aim of this study is to assess in vitro the ability of single or
sequentially combined exposure to very low doses of SKA IL-4
and IL-12 (10-8 µg/ml) to enhance DCs’ antigen presentation
capacity compared to the normally administered doses of
recombinant human (rh) IL-4 and IL-12.
© Dipartimento Scientifico Guna S.p.a.
EX VIVO STUDY
16 subjects
(10 males, 6 females; mean age 73; range 57-83)
With colon carcinoma
9 primary colon
carcinoma
16
2 Dukes A
7 Dukes B 1-2
7 metastatic colon
carcinoma
12 subjects
(7 males, 5 females; mean age 64; range 37-85)
Healthy donors
Dukes C 1-2
with lymph
node
metastasis
EX VIVO STUDY
INCLUSION CRITERIA
• Subjects affected by primary colon carcinoma needing surgery without
pre-surgery evidence of distance metatstasis a
PREPARATIONS
• SKA-IL-4 (10 fg/ml) GUNA Laboratories
• SKA-IL-12 (10 fg/ml) GUNA Laboratories
• rhIL-4 (1ng/ml) Pepro Tech Inc.
• rhIL-12 (1ng/ml) Pepro Tech Inc.
RESULTS
(A) MoDC (monocyte derived denditic cells) allostimulatory
activity of colon carcinoma patients (n = 16) versus
healthy donors (n = 12). (B) MoDC allostimulatory
activity of nonmetastatic (n = 9) versus metastatic (n
= 7) colon carcinoma patients. MoDCs, generated by
culturing PB CD14+ cells from tumor and normal
subjects in the presence of rhGM-CSF and rhIL-4 for
6 days, were incubated with 1 × 105 allogeneic naïve
CD4+ T cells at 1:40, 1:20, and 1:10 ratios for 5 days
followed by a 6-hour pulse of 3H-TdR. Results are
expressed as mean ± SE cpm of triplicate cocultures. Statistical significance was determined
using one-way ANOVA.
Translational Oncology 2015 8, 327-338DOI: (10.1016/j.tranon.2015.06.005)
RESULTS*
RECOMBINANT HIGH DOSE
LOW DOSE SKA
Effects of standard-dose rhIL-4 and/or rhIL-12 and lowdose SKA-IL-4 and/or SKA-IL-12 on APC activity in MLR of
MoDCs
from nonmetastatic colon carcinoma patients (n =
.
6) and from metastatic colon carcinoma patients (n = 7).
MoDCs were untreated or pretreated with SKA-IL-4 (48
hours) and SKA-IL-12 (24 hours) as single agents or
sequentially in parallel to the rh cytokines, and subjected
to MLR with allogeneic naïve T cells in different MoDC-toT cell ratios. The figure shows the mean percentages ± SE
of 3H-TdR incorporation in cpm. Statistical significance was
determined using one-way ANOVA.
Rh/SKA cytokine pretreated nonmetastatic/metastatic
colon
carcinoma
MoDCs
versus
untreated
nonmetastatic/metastatic colon carcinoma MoDCs: *P <
.05 and **P < .01.
Nonmetastatic colon carcinoma MoDCs versus metastatic
colon carcinoma MoDCs: ●●P < .01 and ●●●P < .0001.
Translational Oncology 2015 8, 327-338DOI: (10.1016/j.tranon.2015.06.005)
GUNA IMMUNITY SYSTEM ACTIVATION IN CANCER
IL-12 (10 fg/ml SKA)
IFN-g (10 fg/ml SKA)
IL-4 (10 fg/ml SKA)
IL-12 (10 fg/ml SKA)
FOR FURTHER INFORMATION
Biomedico
www.biomedico.ie
[email protected]
Facebook: biomedico
Tel: 0873275484 - 0874102519
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