Heterochronic parabiosis: the promise of pro- and anti-geronic factors Joseph M. Castellano, Ph.D. Stanford University School of Medicine, Wyss-Coray Laboratory 2016 OAIC Annual Meeting, April 19, 2016
Aging is a major risk factor for disease • Normal brain aging alterations: – Cognitive – Cellular – Molecular • Aging is major risk factor for many neurological disorders • Aged population projected to double by 2030 Young (31 y) Old (88 y)
Young Old Positive factors Positive factors Systemic environment
Young blood regulates aging processes in diverse tissues Conboy et al., 2005 Sinha et al., 2014 ? Castellano et al., JAMA Neurol , 2015 (for review)
Young blood regulates aging processes in diverse tissues Conboy et al., 2005 Sinha et al., 2014 ? Conboy et al., 2005 Castellano et al., JAMA Neurol , 2015 (for review)
Young blood regulates aging processes in diverse tissues Conboy et al., 2005 Sinha et al., 2014 ? Conboy et al., 2005 Castellano et al., JAMA Neurol , 2015 Salpeter et al., 2013 (for review)
Young blood regulates aging processes in diverse tissues Conboy et al., 2005 Sinha et al., 2014 ? Loffredo et al., 2013 Conboy et al., 2005 Castellano et al., JAMA Neurol , 2015 Salpeter et al., 2013 (for review)
Young blood regulates aging processes in diverse tissues Conboy et al., 2005 Baht et al., 2015 Sinha et al., 2014 ? Loffredo et al., 2013 Conboy et al., 2005 Castellano et al., JAMA Neurol , 2015 Salpeter et al., 2013 (for review)
Young blood regulates aging processes in diverse tissues Villeda et al., 2011 Villeda et al., 2014 Katsimpardi et al., 2014 Conboy et al., 2005 Baht et al., 2015 Sinha et al., 2014 ? Loffredo et al., 2013 Conboy et al., 2005 Castellano et al., JAMA Neurol , 2015 Salpeter et al., 2013 (for review)
Young blood regulates aging processes in diverse tissues Villeda et al., 2011 Villeda et al., 2014 Katsimpardi et al., 2014 Conboy et al., 2005 Baht et al., 2015 Sinha et al., 2014 ? Loffredo et al., 2013 Conboy et al., 2005 Castellano et al., JAMA Neurol , 2015 Salpeter et al., 2013 (for review)
Adult neurogenesis occurs in lateral ventricles and the hippocampus (DG)
Signals in the systemic environment regulate neurogenesis in dentate gyrus Villeda et al., 2011
Pro-aging factor CCL11 increases with age and impairs learning/memory Villeda et al., 2011
Pro-aging factor CCL11 increases with age and impairs learning/memory B2M also revealed to be anti-neurogenic, pro-aging factor Smith et al., 2015, Nat Med Villeda et al., 2011
Blood-borne factors that promote plasticity and/or cognition e.g., GDF11, many others
Exposure to young blood revitalizes the aged brain Plasticity gene networks Rejuvenation Dendritic spines/LTP Young Neurogenesis blood plasma Microgliosis Learning/memory Villeda et al., Nat Med, 2014 and unpublished
Strategy for the identification of rejuvenating factors Heterochronic Parabiosis Mouse Aging Human Donors Cellular/molecular changes Cognitive assessment
Strategy for the identification of rejuvenating factors Umbilical cord ~ 20 years Heterochronic Parabiosis ~ 65 years Mouse Aging Human Donors Cellular/molecular changes Cognitive assessment
Protein microarray to assess relative plasma protein expression 150 µ m spot Plasma protein factors Cytokines Chemokines Growth factors Neurotrophins Hormone-like proteins Acute-phase proteins Complement factors Secreted receptors Sample array w/ differential plasma protein signals
Protein microarray reveals many elevated factors in cord plasma Cord Young Old Low High Secreted Plasma Factors Castellano et al., in revision
Protein microarray reveals many elevated factors in cord plasma Cord Young Old Low High Secreted Plasma Factors Castellano et al., in revision
Administration of human plasma in “NSG” mice Heterochronic Parabiosis Changes Mouse Human
Administration of human plasma in “NSG” mice Heterochronic Parabiosis Changes Mouse Human NOD.Cg- Prkdc scid Il2rg tm1Wjl /SzJ aka “NOD Scid Gamma” (NSG)
Age-dependent changes in immediate early gene marker c-Fos Castellano et al., in revision
NSG mice exhibit other age-dependent hippocampal pathology Castellano et al., in revision
NSG mice exhibit age-dependent learning and memory deficits Castellano et al., in revision
NSG mice exhibit age-dependent learning and memory deficits Castellano et al., in revision
Administration of human plasma in aged NSG mice Microarray/qPCR/IHC 0 14 Days Human plasma Aged NSG donors Behavior
Human plasma treatment results in distinct gene profiles in hippocampi Castellano et al., in revision
Human plasma treatment results in distinct gene profiles in hippocampi Ontology-based activation prediction -Long-term memory -Long-term potentiation -neuritogenesis Plasticity upregulation by qPCR -c-Fos, Egr1, junb, camk2a, etc. Castellano et al., in revision
Cord plasma increases c-Fos+ cell number in aged but not young dentate gyrus Aged NSG Castellano et al., in revision
Cord plasma increases c-Fos+ cell number in aged but not young dentate gyrus Aged NSG Young NSG Castellano et al., in revision
Cord PLM activates DG granule neurons (excitatory) Castellano et al., in revision
Treatment with cord plasma enhances LTP in dentate gyrus RE SE Castellano et al., in revision
LTP is unaffected following young or elderly human plasma treatment Castellano et al., in revision
Treatment with cord plasma improves learning and memory performance Castellano et al., in revision
Strategy for the identification of rejuvenating factors Heterochronic Parabiosis Umbilical cord Mouse Aging Human Aging plasma Cellular/molecular changes Cognitive assessment
Strategy for the identification of rejuvenating factors Heterochronic Parabiosis Mouse Aging Human Aging Cellular/molecular changes Cognitive assessment
In vivo “screen” of potential rejuvenating factors in aged WT mice Putative Factors or vehicle i.p. 0 2 4 6 10 Days Castellano et al., in revision
Plasma TIMP2 is reduced very early in life in blood and brain Human Mouse Castellano et al., in revision
Systemic supplementation with TIMP2 enhances synaptic plasticity in aged mice rTIMP2 or vehicle i.p. 0 2 4 6 8 10 12 14 Days Castellano et al., in revision
Systemic treatment with TIMP2 improves learning and memory in aged WT mice Castellano et al., in revision
Systemic TIMP2 is necessary for spatial memory Training Day 1 Novel Location Day 2 Castellano et al., in revision
Translational potential of blood-borne proteins • TIMP2 – 21-24 kDa; non-glycosylated – Limits tumor angiogenesis and plays role in tissue remodeling – Broad MMP inhibitor Marimastat proceeded to Phase III trials (discontinued) Ongoing plasma trials related to CNS • – AMBAR (Alzheimer’s Management by Albumin Replacement - Grifols) – PLASMA ( PL asma for A lzheimer’s S y M ptom A melioration)
Conclusions • Blood factors can regulate aging process in diverse tissues – Developmental-stage human plasma (UC) improves neuronal and cognitive function – Young plasma proteins (TIMP2) restore or improve function of aged hippocampus – Use of NSG mouse model allows for identification of relevant blood-borne proteins with possible translational implications
Acknowledgements Wyss-Coray Lab Stanford Tony Wyss-Coray Tom Rando Kira I. Mosher Martin Angst Rachelle Abbey Daniela Berdnik Funding Sources Jadon Shen -NIH/NIA -Jane Coffin Childs Postdoctoral Fellowship/Simons Foundation Xie Lab/AfaSci -Stanford Child Health Research Institute Simon Xie Postdoctoral Fellowship Bende Zou -Donor’s Cure Foundation, CCAD
Young blood alters brain via unknown mechanisms/factors Synaptic plasticity Oligodendrocyte activity Neurogenesis Cell-to-cell Signaling? Transport? Diffusion? Secondary signals? Disrupted BBB? Youthful Youth-associated Stimulated systemic leukocytes? vascular factors endothelium Castellano et al., JAMA Neurol , 2015
Human plasma does not alter the number of neuroblasts in dentate gyrus of NSG mice Castellano et al., in revision
rTIMP2 treatment does not appear to alter DCX+ cell number in aged DG Castellano et al., in revision
TIMP2 expression in plasma and hippocampus declines gradually with age in mice Castellano et al., in revision
Blood-borne factor GDF11 mediates SVZ neurogenesis rejuvenation Katsimpardi et al., 2014
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