Sirtuins and NAD

Joseph BaurPerelman School of MedicineUniversity of Pennsylvania

DoD State of the Science SymposiumOctober 25, 2019

What causes aging?

Can we slow it down?

50 71

49 69?

Caloric Restriction Increases Rodent Lifespan

Barger et al. Exp Gerontol. 38:1343 (2003)

ALCR

Female Mice

Sun et al. Clin. Diagn. Lab. Immunol. 2001, 8(5):1003.

Paraquat Injection

…and Stress Resistance

Will targeting longevity pathways conferstress-resistance like CR?

Carrie Sims, MD, PhD

Associate Professor of Surgery

Hospital of the University of Pennsylvania Presbyterian Medical Center of Philadelphia

Hemorrhagic Shock

Brohi et al. Intensive Care Med, 2019.

Mortality for Trauma Patients Activating a Hemorrhage Protocol and Requiring

Blood Transfusion

How Does CR Work?

Energy Intake

Body Size?

Adiposity?

Metabolic Rate?

Oxidative Stress?

Insulin Signaling?

Chronic Inflammation?

GH/IGF1 Signaling?

Ingestion of AGEs?

Hormesis?

mTOR Signaling?AMPK Activity?

NAD?

…and can we mimic it with drugs?

Sirtuins ?

Mimicking CR-Induced Stress-Resistance

SIRT1

Resveratrol

Resistance to Trauma, Hemorrhagic Shock, Sepsis, Chemotherapy???

?

Rat Hemorrhagic Shock Model

Anesthetized:LinesLaparatomy

30 min stabilizationEmerge from anesthesia

Begin hemorrhageto MAP 40

mmHg

Severe Shock

Resuscitation over 60 min with4X shed volume in LR

- or –4x shed volume in LR + RSV

60 min

60 R 18 hr

18 hours

Decompensation: Incremental Infusion

Req’dto maintain MAP

Physiologic Reserve

Resveratrol Ameliorates Hyperglycemia During Resuscitation from Hemorrhagic Shock

Hyperglycemia Lactate

RSV

Wang et al, J Trauma Acute Care Surg. 2014Wang et al, Shock. 2015

Resveratrol Ameliorates Hyperglycemia During Resuscitation from Hemorrhagic Shock

Wang et al, J Trauma Acute Care Surg. 2014Wang et al, Shock. 2015

RSV RSV

Resveratrol Ameliorates Oxidative Stress Induced by Hemorrhagic Shock

Kidney ROS Liver ROS

RSV RSV RSV RSV

Wang et al, J Trauma Acute Care Surg. 2014Wang et al, Shock. 2015

Resveratrol Ameliorates Mitochondrial Dysfunction Induced by Hemorrhagic Shock

Mitochondrial Respiration

Wang et al, J Trauma Acute Care Surg. 2014Wang et al, Shock. 2015

RSV

Mimicking CR-Induced Stress-Resistance

SIRT1

Resveratrol

Resistance to Trauma, Hemorrhagic Shock, Sepsis, Chemotherapy???

??

Nicotinamidemononucleotide

(NMN)

NAD

?

NMN Increases Tissue NAD Levels

Sims et al, JCI Insight. 2018

NMN Ameliorates Hyperglycemia During Resuscitation from Hemorrhagic Shock

Sims et al, JCI Insight. 2018

NMN Ameliorates Mitochondrial Dysfunction Induced by Hemorrhagic Shock

Mitochondrial Respiration

Sims et al, JCI Insight. 2018

Palmitoyl Carntine

Pyruvate/Glutamate

/MalateSubstrate:

pmol

O2/

s/m

g

NMN Improves Survival Following Hemorrhagic Shock

Sims et al, JCI Insight, 2018

Summary of Hemorrhagic Shock Data

Both resveratrol and NMN are protective, suggesting a role for SIRT1: Controlled hyperglycemia and reduced lactate Improved mitochondrial function Improved survival (only NMN tested)

Mechanistic studies using genetic tools have been limited by the need to use rats

Hemorrhagic Shock in Mice

base CI M+P CI M+P +G CII CIV

Kidney Mitochondrial Respiration

WH sham

WT HS

* *

*

Qingwei Chu

SIRT1 KO?

Resveratrol Improves Survival Following Septic Shock

Sims et al, unpublished

Cecal Ligation/Puncture

SIRT1 KO Worsens Survival Following Septic Shock

Sims et al, unpublished

Loss of SIRT1 is Pro-Inflammatory

Sims et al, unpublished

IL6 in Circulation IL6 mRNA in Bone Marrow-Derived Macrophages

Macrophage-Specific Loss of SIRT1 is Pro-Inflammatory

Sims et al, unpublished

IL6 in Circulation Survival

LysM Cre

Deletion of SIRT1 in macrophages onlyX

SIRT1 Flox

Septic Shock

Despite some conflicting pharmacological data in the literature, SIRT1 appears to be beneficial in septic shock

SIRT1 acts at least partially through anti-inflammatory mechanisms in macrophages

Are other sirtuins important? Other cell types?

SIRT3

Mitochondrial sirtuin involved in oxidative stress resistance

Sims et al, unpublished

SIRT3 Does Not Protect Mitochondrial from Septic Shock

Mitochondrial Respiration in Kidneys of Survivors 5 Days After Septic Shock

Sims et al, unpublished

Loss of SIRT3 is Pro-Inflammatory

Sims et al, unpublished

IL6 in Circulation IL6 mRNA in Bone Marrow-Derived Macrophages

Summary

Multiple sirtuins, including SIRT1 and SIRT3, are likely to play roles in critical injuries

A major role of sirtuins is to suppress the inflammatory response in macrophages

Timing and cell-type specificity of interventions will likely be critical…genetic models are extremely useful to tease these details out

Translation

Resveratrol and NMN could be moved into larger animal models after a few additional studies to address timing and optimal dosing.

There is potential to greatly improve the efficacy of these interventions by using genetic tools to understand the mechanisms.

Applying to NIH and DoD opportunities (Congressionally Directed Medical Research Program)

AcknowledgmentsBaur Lab

Karthi ChellappaSarmistha MukherjeeTim LuongoLauren PaolellaJim DavisQingwei ChuXiaoyan YinJames MoYashu BhatCaroline Perry

Past MembersFunding

NIDDKNIAPenn Center for Musculoskeletal Disorders Pilot Award

Will QuinnCarrie SimsDavid FrederickCole PurdyDenice DeFelisePaul BotolinYuxia GuanTony Davila

Hao WangCarrie Sims

Qingwei ChuAnna Garcia-

Whitlock

Resveratrol and Rapamycin Prolong Survival in a Sepsis Model

Holthoff et al. Kidney International. 2012, 81:370

Resv

Ctrl

Days

Yen et al. Surgery. 2013, 153:689