Gladstone Institute

• Ken Nakamura (a neurologist)
  • Nakamura’s lab has developed tools to monitor ATP production and study metabolism in normal and diseased neurons,
  • They have also developed cutting-edge approaches to track the fates of individual mitochondria in neurons over time,
  • His age-related mitochondrial dysfunction ((with PhRET, CompBio, ARN);
• My proposal
  • (Mendelsohn, 2018 #1380) to identify genetic defects that will be responsive to energy-based therapies.!
    • Low-ATP hits: 144 FRET-increasing hits→ 797개 remaining in the glycolytic condition,→ 43 hits (protecting ATP in glycolysis also decreasing ATP in the respiratory condition)
    • 특히, Genes that decrease ATP when knocked down (Knocking down of COX11, KPNB1, PDSS1, PDSS2, ATP5MPL significantly decreased maximal respiration (after FCCP).) fig4, Mendelsohn 2018)
  • The link between mitochondrial/ATP deficit and neuronal cell death:
    • Quantitative correlation between mitochondrial/ATP reduction (or any other mitochondrial parameters) and neuronal cell death
    • Will ATP restoration slow down neuronal cell death?
    • Deep phenotyping of the cells with mitochondrial deficit synapse, aSyn
  • The link between mitochondrial dysfunction and aSyn
    • Will mitochondria/ATP restoration slow down aSyn spreading?

In sPD	?aSyn?	↓ mito function	aSyn?	↓ ATP	NDGN	Motor symptom	Detailed analysis	In vivo
		What readouts?			Why SN?		Mitochondrial detailed analysis	aSyn PFF
							Transcriptional analysis	Line61
Tx		How to identify?					Metabolomic anlysis

• Original proposal

	gene			↓ ATP
				aSyn
				survival
				5 different PD subtypie
correction	(Mendelsohn, 2018 #1380) 이렇게 identify인 gene을 KD 한 후→ CoQ10 주니까→ ATP restored

• Takeda’s proposal (AIM0)

				Aging	mito
				aSyn
correction

• Cells

Young	iN		iPSC	sPDD iN	sPDD iPSC
aged	iN		iPSC

• Questions

Why PDD?
Budget?

Glutamate Imaging

		Example
Chemical Exchange Saturation Transfer (CEST)	MR molecular imaging approach that utilizes a frequency selective radiofrequency (RF) irradiation pulse on particular exchangeable protons (e.g., hydroxyls, amides, and amines), thus resulting in attenuated water signals that can be measured via the loss of water signal intensity to indirectly characterize the microenvironment of the solution (Ward et al., 2000; Li et al., 2017) amine protons on Glu that show a chemical shift of 3.0 ppm away from bulk water (0 ppm) can be measured In MPTP model: (Bagga, 2018 #1933) Repeated measurements in five mice demonstrated an intra-animal coefficient of variation (CV) of GluCEST signal to be 2.3 ±1.3% and inter-animal CV of GluCEST to be 3.3 ± 0.3%. Mice were treated with MPTP to create a localized striatal elevation of glutamate. We found an elevation in the GluCEST contrast of the striatum following MPTP treatment (Control: 23.3 ± 0.8%, n = 16; MPTP: 26.2 ± 0.8%, n = 19; p ≤ 0.001). Figure 4. GluCEST MRI and ¹H MRS in the striatum. (a) GluCEST contrast was found to be higher in the striatum of the MPTP group (26.2 ± 0.8%, n = 19) compared to the control group (23.3 ± 1.0%, n = 16; p ≤ 0.001). (b) Glutamate measured via ¹H MRS in the striatum was also found to be higher in the MPTP group (control: 12.3 ± 1.0 mM, n = 16; MPTP: 14.4 ± 1.1 mM, n = 19; p ≤ 0.005). the positive association between glutamate concentration measured via 1H MRS and GluCEST signal was used to estimate background contribution to the measured GluCEST. The contribution of signal from non-glutamate sources was found to be ~28% of the total GluCEST. Immunohistochemical analysis of the brain showed co-localization of glutamate with GFAP in the striatum. Figure 5. Correlation between GluCEST contrast and ¹H MRS. Plot showing the mean values of GluCEST contrast and glutamate concentration in the striatum for the control and MPTP treated mice. All data are presented as Mean ± SD.	BRI, Niigata Univ (AQP4 potentiator, PS19 mice)
11C-ABP688, 4 MRS	A PET tracer with highly specific binding to the metabotropic glutamate receptor 5 (mGluR5)	Ad/pd 2023, Christopher Doppler: ↑ in PD+RBD, E.J.

Goals

FY22

Experimental Medicine – KPIs for 2022

Mission: Experimental medicine is the use of innovative measurements, models and designs in studying human subjects and samples for establishing proof of mechanism and concept of new drugs, and for exploring the potential for market differentiation for drug candidates

Disease Area Translational strategy	Target validation	Translational resources
For all DAUs, bring together with the biology and clinical lead a disease area Translational Medicine Plan (TMP) for each priority indication or target draft by June Identify key breakthrough translational gaps for each DAU and prioritize gaps Work with MBM, Ephys, Human Biology, PTS, and DSI (as needed) on delivering on at least 80% of breakthrough biomarkers Upcoming portfolio milestones by DA: PEs (DMPK, NRLP3, Parkin, etc.) LGEs (PMP22, etc.)	Work with DAUs and BTUs to prioritize key targets for validation and include target validation plans within the DA-TMPs by June Work with Biology, Clin Sci, NSTM, Comp Bio, and Stats on execution Support cross-DDU target discussion from a translational perspective for new target discussion Obtain samples and tissue for 80% of priority target assessment	For all DAUs, map community efforts, fluid and sample access and patient data each priority indication or target by June Work with biology, Clin Sci, NSTM and others on access to all (100%) high priority resources Work with NSTM LT and PQIB on budget requests and prioritization

TIMELINE: target July 8th to have your individual draft goals entered, → which the NSTM LT will review and prioritize at our next meeting on Mon July 11th

• Align with BOB
• Consider relocation

NSTM Pre-CS Key Objectives in PD:
High priority NLRP3 human data for TV (PD, AD)

• Obtain samples and tissue for 80% of priority target assessment
  
• Align project TV and TM plans across the DAU

DA TMP (June)?

Identify/prioritize key breakthrough translational gaps → delivering on at least 80% of breakthrough biomarkers (Ceri: “Something we can measure that now provides us with the ability to make a progression/termination decision that we (and our competitors) did not have the ability to do before)
Upcoming portflio milestones : NLRP3
Translational resources: - map community efforts, fluid and sample access and patient

data by June
[my summary]

• NLRP3 TV with human data (PD, AD)
• Project milestone: NLRP3
• Translational resources: Obtain/map human samples and tissues and access
• Identify/prioritize/deliver key breakthrough translational gaps and biomarkers
• Align project TV and TM plans across the DAU: Using NLRP2 and SNCA as building block to develop DAU alignment (process, Target validation framework and “minimal package for PD”)

To add:

• NLRP3 segmentation strategy?
• Parkin GT : natural Hx study?

1. Your FY22 goals
2. Your Individual development plan (template attached)

• Develop and execute translational plans of NLRP3, PRKN GT and SNCA BTV to adress the key project needs
• Develop and execute on the translational strategy for Parkinson’s disease as a disease area – Identify and target key translational gaps and strategy
• Resolves key disease area translational risks by accessing external translational resources and platforms:
  • Patient-derived sample access in PD to support target valiation and biomarker validation

Uncertain Spans

location	transcription	uncertainty
Original proposal / 5 different PD subtypie	reads as written; subtypie typography preserved verbatim.	source typography.
Takeda's proposal (AIM0) heading	reads with terminal 0 glyph; preserved verbatim.	low confidence on O vs 0.
Mendelsohn 2018 / 797개	the count reads 797 followed by Korean 개; preserved verbatim.	low confidence on the digit boundary.
Experimental Medicine slide / NRLP3	the slide bullet reads NRLP3 (note NRLP ordering), not NLRP3; preserved verbatim.	source typography preserved.