Le potenziali applicazionidella NANOMEDICINA alle

malattie neurodegenerative

Dr. Francesca Re

School of Medicine and Surgery - Nanomedicine Center Biochemistry Unit

University of Milano-Bicocca, Monza (IT)

30 Marzo 2018

Uno sguardo al futuro…

Siamo nel 2083. Anna ha 110 anni, è ancora ingamba, ma inciampa in un tappeto e caderompendosi il femore; resta distesa adaspettare e nel giro di una decina di minutipotrà rialzarsi in piedi poiché il femore siè auto-riparato. Come è potuto accadere?Dall’anno 2070 gli ultracentenari assumononano-particelle autoadesive che vanno arimpiazzare una parte dei costituenti delleossa.

Non stiamo parlando di fantascienza, ma dell’ultima frontiera della medicina, che fa già oggi uso di minuscoli dispositivi di qualche miliardesimo di metro programmabili, intelligenti, e capaci di svolgere diversi compiti contemporaneamente per curare o diagnosticare malattie.

Qual è il presente della piccola scienza di domani?

Un semplice esempio per capire le potenzialità delle nanotecnologie:

la natura sfrutta da moltissimo tempo la nanostrutturazione della materia vivente per ottenere proprietà assolutamente “nuove” fino a poco tempo fa ritenute irraggiungibili

copiando la natura (“materiali bioispirati”) anche l’uomo sta avvicinandosi a queste “nuove” proprietà

Il geco cammina sul soffitto grazie alla nanostrutturazione dei suoi polpastrelli

La sommatoria di un elevatissimo numero di piccolissime forze di attrazione supera la forza gravità

L’uomo è riuscito a riprodurre questa nanostrutturazione:

La Nanomedicina è definita come la applicazione delle nanotecnologie

alla salute umana. Sfruttando le migliorate e spesso nuove proprietà

fisiche, chimiche e biologiche dei nanomateriali.

[US-National Institute of Health Roadmap for Medical Research in Nanomedicine]

Definition of NANOMEDICINE

All-in-one nanodevice: NANOPARTICLESSmart nanostructures for diagnosis and therapy

with a size of 1-100 nanometers

[US-National Institute of Health Roadmap for Medical Research in Nanomedicine]

Applications of Nanomedicine

o Diagnosis

o Therapy

o Regenerative medicine

Advantages:

o Drug targeting

o Lesser drug is necessary

o Lesser side effects

o Protection of drugs from degradation

longer half-life

o Multi-functionalization

Multi-functionalizationTHERANOSTIC

Nanoparticles features make them ideal for therapy and diagnosis of different diseases,

including disorders of the central nervous system

BLOOD

BRAIN Blood-Brain Barrier (BBB)

Physical and biochemical barrier

that regulates the flow of molecules

between blood and brain

Alzheimer’s Disease (AD)

o Neurodegenerative disease that culminates in total

loss of cognition and executive functions

o Worldwide, nearly 30 million people have AD

o Only 1-in-4 people with AD have been diagnosed

o Sporadic (majority). Only 5 % cases are familial forms

http://www.alz.org/

No cure for AlzheimerCurrently available drugs may help

to prevent some symptoms

Hallmarks of Alzheimer’s Disease

o Plaques (Aβ peptide)

o Tangles (hyperphosphorylated Tau)

Nanoparticles for therapy and diagnosis of Alzheimer’s disease

Design and development of advanced nanomedicines to overcome biological barriers and to treat severe

mApoE-Functionalized Lipidic -and Polymeric- Nanocomposite for Human Glioblastoma Imaging and Treatment

Smart Nanoparticles For Boosted Drug Brain Targeting

Our approach to Alzheimer’s Disease

Development of a Novel Multicellular In Vitro Model of Alzheimer’s disease-like Blood-Brain Barrier

Sphingomyelin/cholesterol (1:1 M/M)5 mol% phosphatidic acid2.5 mol% DSPE-PEG-maleimide

PEG

mApoE

Liposomes (mApoE-PA-LIP) able to (in vitro)

bind Ab with high affinity

disaggregate Ab assemblies

slow down Ab aggregation

overcome a Blood brain barrier model

be biocompatible

Diameter 100-150 nm

ARE THESE NP SUITABLE FOR TREATING

ALZHEIMER DISEASE IN VIVO ?

[Bana L. et al., Nanomedicine 2014]

mApoE-LIP reach intact mouse brain after systemic administration

Confocal microscopy (hippocampal regions)

BODIPYFL-sphingomyelin

Rhodamine B

[Balducci C. et al., J. Neurosci. 2014][Bana L. et al., Nanomed. 2014]

APP/PS1

DOUBLE TRANSGENIC

Animal model

10-12 MONTHS OF AGE:

Evident amyloid plaques Evident cognitive deficits

[Garcia-Alloza M et al., Neurobiol Dis 2006]

APPswe PSEN1dE9

5 MONTHS OF AGE:

Initial Aβ deposition Absence of cognitive deficits

Model of PRE-SYMPTOMATIC AD

Model of SEVERE AD

Proof-of-concept in vivo

• Therapeutic treatment on SEVERE AD Tg mice

APP/PS1 of 10 months of age

Intraperitoneal injection

100mL of [40mM] mApoE-PA-LIP

(total lipids = 2.5 mg/injection)

3 injection/week for 3 weeks

Translational issues:• Irreversible brain damages present at severe AD stage

in humans• Failure of clinical trials focused on mild to moderate

stages of AD

Aβ accumulation in the brain begins ~10–20 years prior to the onset of clinically detectable symptoms

This early stage is considered the most promising time frame to start with disease-modifying therapies

APP/PS1

DOUBLE TRANSGENIC

Animal model

10-12 MONTHS OF AGE:

Evident amyloid plaques Evident cognitive deficits

[Garcia-Alloza M et al., Neurobiol Dis 2006]

APPswe PSEN1dE9

5 MONTHS OF AGE:

Initial Aβ deposition Absence of cognitive deficits

Model of PRE-SYMPTOMATIC AD

Model of SEVERE AD

Proof-of-concept in vivo

• Preventive treatment on PRE-SYMPTOMATIC AD Tg mice

APP/PS1 of 5 months of ageIntraperitoneal injection100mL of [40mM] mApoE-PA-LIP (total lipids = 2.5 mg/injection) 1 injection/week for 7 months

At the end of each treatment mice were analyzed for:

o Memory impairment by novel object recognition test (NORT)

o Brain Ab content by immunohistochemical, biochemical and

imaging assays

o Amount of Ab in liver and spleen by ELISA assay

AFTER ACUTE TREATMENT WITH mApoE-PA-LIP

o Treated mice significantly recovered their long-term

recognition memory

[Balducci C. et al., J. Neurosci. 2014]

AFTER ACUTE TREATMENT WITH mApoE-PA-LIP

o Strong reduction (-35%) of amyloid plaques and Ab burden in

the brain of treated mice

[Balducci C. et al., J. Neurosci. 2014]

PREVENTIVE TREATMENT WITH mApoE-PA-LIP

o Prevents the onset of memory impairment and the occurrence

of AD-like cerebral abnormalities

[Mancini S. et al. J Cont Rel 2017]

PREVENTIVE TREATMENT WITH mApoE-PA-LIP

o Strongly reduces (-30%) the amyloid plaques deposition in the

brain

[Mancini S. et al. J Cont Rel 2017]

AFTER TREATMENTS WITH mApoE-PA-LIP

o Increase of Ab in the liver and spleen

[Mancini S. et al. J Cont Rel 2017]

[Balducci C. et al., J. Neurosci. 2014]

LIVER SPLEEN

PEG

mApoE

PA

Plaques

BBB crossing

Soluble Ab

assemblies

BLOOD

BRAIN

Liverspleen

MoA of mApoE-PA-LIP

We developed a nanomedicine able to

o target and reduce brain Ab

o hinder the memory impairment

mApoE-PA-lip (Amyposomes®) are promising

nanodevices for prevention and treatment of AD

The aim of the NewCo is to bring Amyposomes® to the P-o-C clinical phases in humans

School of Medicine Prof. Massimo MasseriniPoC in animal models Human samplesS. Mancini C. FerrareseG. Sancini C. ZoiaV. Zambelli E. ContiPoC in vitroI.RivoltaFunctionalizationM.GregoriS.SesanaBiocompatibilityE. CazzanigaA. CoxCharacterizationF.MantegazzaD.Salerno

Dept. Biotechnologyand BioscienceChemical synthesisF.NicotraB. La FerlaC. Airoldi

Claudia BalducciGianluigi ForloniMario SalmonaMarco GobbiPaolo Bigini

Turku PET Centre

Juha RinneMerjia HapaarantaAnniina Sleman

F. WandosellA. Ordonez-Gutierrez

Thanks to …

available to collaborations

…Ritornando

al futuro…

NANOROBOTS: FACT OR FICTION?

Thanks for your attention

mApoE-PA-LIP effects were maintained 3 monthsafter treatment discontinuation

LRP-1 RAGE