Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
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. © Dipartimento Scientifico Guna S.p.a. 14 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 16 © Dipartimento Scientifico Guna S.p.a. 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 © Dipartimento Scientifico Guna S.p.a. 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 © Dipartimento Scientifico Guna S.p.a. © 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 © Dipartimento Scientifico Guna S.p.a. 37