the use of adxs-her2 in canine...

2015 Dr. Nicola Mason

Nicola J. Mason, B.Vet.Med., Ph.D., Assistant Professor of Medicine and Pathobiology at the

University of Pennsylvania’s School of Veterinary Medicine

The Use of ADXS-HER2 in Canine

Osteosarcoma

102

2015 Dr. Nicola Mason 103

Canine and Human Osteosarcoma

comparable behavior, biology, genetics and therapeutics

H&E HER2/neu

Paoloni et al. BMC Genomics 2009

<< Pr e v N ext >>Fr o m : An n Th o ra c M ed . 20 1 0 Ap r- Ju n; 5 (2) : 6 7 –7 9 .

doi : 10. 4103/ 1817-1737. 62469

Co p yr igh t/ Lice n se ! Re qu e st pe r m issi on to r e use

Figure 19

A chest ra diog ra ph sho wing ca lcif ied metast ases fro m an osteo genic sa rco ma. No te that t he densit y of t he

tumors and the sk eletal tissue s is sim ilar

Im ag es in th is ar ticle

PubM ed C entr al, F igur e 19: A nn T hor ac M ed. 2010 A pr- Jun; 5( ... http: //pr oxy .l ibr ary.upe nn.e du: 2082/ pm c/articles /PMC 2883201/ ...

1 of 3 2/3/ 13 10: 07 P M


ADXS-HER2: Canine Osteosarcoma

Phase 1 n=18

Canine Osteosarcoma

University of Pennsylvania’s School of Veterinary

Medicine

Canine and human osteosarcoma are not

distinguishable by global gene expression analysis (Paolini M., BMC Genomics, 2009)

Canine and human HER2 have high degrees of

homology

Under review at USDA for veterinary use

Currently being evaluated in combination with RT

Objectives

Determine MTD of ADXS-HER2

Determine whether administration is safe and

capable of eliciting tumor-specific immunity

Determine if ADXS-HER2 can prevent metastatic

disease and prolong overall survival

ADXS +

Standard Tx

Carboplatin

q21 days x 4

ADXS

Treatment

Diagnosis &

Amputation

Screening

visit ADXS ADXS

104


No Evidence of Long or Short Term

Cardiotoxicity Following AT-014 Administration

Cardiac Troponin I Fractional shortening

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

2.0

2.2

2.4

2.6

2.8

0 4 8 12 16 20 24 28 32 36 40 44 48 52 56 60 64 68 72 76 80 84 88 92 96100104108112

CardiacTroponinILevel(ng/mL)

Weeks post first vaccination

2 x 10^8

5 x 10^8

1 x 10^9

3 x 10^9

0

10

20

30

40

50

60

70

0 4 8 12 16 20 24 28 32 36 40 44 48 52 56 60 64 68 72 76 80 84 88 92 96 100 104 108 112

Fra

cti

on

al S

ho

rten

ing

%


2 x 10^8

5 x 10^8

1 x 10^9

3 x 10^9

0

10

20

30

40

50

60

70

0 4 8 12 16 20 24 28 32 36 40 44 48 52 56 60 64 68 72 76 80 84 88 92 96 100 104 108 112

Fra

cti

on

al S

ho

rten

ing

%


2 x 10^8

5 x 10^8

1 x 10^9

3 x 10^9

No Evidence of Long or Short Term Cardiotoxicity

Following ADXS-HER2 Administration

105

Weeks post first dose Weeks post first dose


Treatment Related Adverse Events

# Patients with Treatment Related Adverse Events

(All toxicities reported are Grade 1)

ADXS-HER2 dose 1x108 5x108 1x109 3x109 Total

Number of dogs recruited N=3 N=3 N=9 N=3 N=18

General Disorders

Pyrexia (>103) 2 1 5 2 10

Fatigue 1 1 7 2 11

GI Disorders

Vomiting 2 1 8 1 12

Nausea 2 1 9 2 14

Cardiovascular

Arrhythmias 0 1 1 1 3

Tachycardia 0 0 1 1 2

Hypotension 0 0 0 0 0

Hematological parameters

Thrombocytopenia 0 0 5 0 5

Biochemical parameters (inc)

γ-GT 0 2 0 0 2

Alkaline Phosphatase 1 1 4 1 7

ALT 1 1 1 0 3

AST 1 1 5 1 8

BUN 0 0 0 0 0

CREA 0 0 0 0 0

Cardiac Troponin I 0 0 1 0 1

106


-50

0

50

100

150

200

250

0

50

100

150

200

250

300%

incr

ease

in W

BC

% in

crea

se in

n

eutr

op

hils

Dead Alive Dead Alive

p=0.003 p=0.003

Early Immune Response to ADXS-HER2

May Predict Survival

0

50

100

150

200

250

300

-50

0

50

100

150

200

250

% in

crea

se in

WB

C

% in

crea

se in

n

eutr

op

hils

2e8 5e8 1e9 3e9

Dose2e8 5e8 1e9 3e9

Dose

107


Evaluation of HER2/Neu Specific T Cell

Responses Induced by ADXS-HER2

108

Induction of HER2-specific immune responses appear to correlate with overall survival


3 weeks post carboplatin 6 weeks post carboplatin

9 weeks post carboplatin 10 weeks post carboplatin

ADXS-HER2 Prevents the Development of

Metastatic Pulmonary Disease

6 yr FS Bernese Moutain Dog

109


Solitary metastatic nodule “Normal” lung tissue

Bright light

Near Infra-Red

light

ICG Tracer Identifies Areas of Inflammation

in Right Cranial Lung Lobe

110


10 weeks post carboplatin

Solitary

nodule

Inflammatory

foci

Immune Cells Target Microscopic

Metastatic Disease

Lymphocytic infiltrate is identified around metastatic nodule and in isolated foci

111


10 weeks post carboplatin

CD3 CD3

CD3 CD3

VimentinVimentin

H&E

H&E

H&E

CD3+ T Cell Infiltrates Associated with

Pulmonary Metastatic Disease

112


Preliminary Data: ADXS-HER2 and Overall Survival

Summary of published data using single

agent carboplatin in dogs with OSA

All dogs without gross metastatic

disease at the time of first dosing

Median survival: Control: 316 days

ADXS: Not yet reached

Survival of Data 11:Survival proportions

0 200 400 600 800 10000

20

40

60

80

100

VACCINE

Control

Days from diagnosis

Pe

rce

nt s

urv

iva

l

n=18

n=11

p<0.0001

Survival of Data 11:Survival proportions

0 200 400 600 800 10000

20

40

60

80

100

VACCINE

Control

Days from diagnosis

Pe

rce

nt su

rviv

al ADXS

113


Synergistic Effects of Radiation and Immunotherapy

Immunogenic cell death

Release of tumor antigens

Dendritic cell recruitment and

activation

Release of HMGB1Exposure of calreticulin

Up-regulation of MHC I

Induction of FAS expression

Induced chemokine production

Up-regulation of ICAM-1

Induction of TGF-βAccumulation of Tregs and M2 macrophages

Antigen-specific T cell priming

T cell recruitment

ADXS-cHER2

+

+

-

Kill primary tumor

Kill distant metastatic tumors

Hypothesis: Combination PTRT plus

ADXS-HER2 will augment anti-tumor

immune responses, prevent metastatic

disease and prolong overall survival in

dogs with appendicular OSA

114


Evaluation of Combination Therapy :ADXS-HER2 and Palliative RT in Primary Osteosarcoma – A Pilot Study

Days 0 4 5 7 28 49 70 91 112 133 154

RT1 RT2

CBC,CS,UA

Immune analysis

EKG/Echo/cTnI

Bone biopsy/histopth

CT – limb and thorax

CBC,CS,UA

Immune analysis

EKG/Echo/cTnI

Bone aspirate/cytology


CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



V#1 V#2 V#3 V#4 V#5 V#6 V#7 V#8

Lameness score; Pain questionnaire

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

EUTHANASIA

CBC,CS,UA

Immune analysis

cTnI

Necropsy

Thoracic radiographs

Limb radiographs

Thoracic radiographs

Limb radiographs

Days 0 4 5 7 28 49 70 91 112 133 154

RT1 RT2

CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



V#1 V#2 V#3 V#4 V#5 V#6 V#7 V#8


Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

EUTHANASIA

CBC,CS,UA

Immune analysis

cTnI

Necropsy

Thoracic radiographs Limb radiographs


Days 0 4 5 7 28 49 70 91 112 133 154

RT1 RT2

CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



V#1 V#2 V#3 V#4 V#5 V#6 V#7 V#8


Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

EUTHANASIA

CBC,CS,UA

Immune analysis

cTnI

Necropsy



Days 0 4 5 7 28 49 70 91 112 133 154

RT1 RT2

CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



CBC,CS,UA

Immune analysis

EKG/Echo/cTnI



V#1 V#2 V#3 V#4 V#5 V#6 V#7 V#8


Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

Immune

analysis

EUTHANASIA

CBC,CS,UA

Immune analysis

cTnI

Necropsy



A A A A A A A A

115


ADXS-HER2 Delayed Progression of Primary Tumor

116

02.22.2012 05.09.201202.01.2012

12.13.2011 2.06.201211.17.2011

7.11.2014 9.23.2014 12.15.2014

Radiation Radiation plus ADXS-HER2


Stable/Improved Function Observed Following PTRT+ ADXS-HER2

117

2.26.2014 11.03.2014

386-003

12.15.20147.11.2014

386-007


Brief Validated Pain Score (owner questionnaire)

118


Preliminary Findings

• ADXS-HER2 may prevent metastatic disease and may prolong survival in dogs with

spontaneous HER2+ OSA when administered in the setting of minimal residual

disease

• ADXS-HER2 appears to break peripheral tolerance to the highly conserved IC1

domain of HER2/Neu

• The magnitude of increase in leucocytes within 24 hours of ADXS-HER2

administration appears to correlate with survival, suggesting that outcome depends in

part upon the ability of the dog’s immune system to respond to the immunotherapy

• ADXS-HER2 at doses up to 3 x 109 cfu has been safely administered to dogs with

spontaneous OSA and has been observed to cause transient, low grade side effects

to date

• Prevention of pulmonary metastatic disease may be in part associated with CD3+

T cell mediated elimination of microscopic metastatic disease in lung which warrants

further investigation

Phase 1 Study: ADXS-HER2 monotherapy post standard of care

Phase 1 Study: Combination palliative radiation and ADXS-HER2 in primary OSA

• Preliminary data that ADXS-HER2 may delay progression of primary tumor,

improve mobility and improve quality of life

119

the use of adxs-her2 in canine...

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