Prospective Evaluation of AI R&D Tool in Adult Glioma and Other Primary Brain Tumours (PEAR-GLIO)
Conditions: Brain Tumor, Glioma, Malignant, Ependymoma, Oligodendroglioma, Glioblastoma
Sponsor: Ourotech, Inc.

 
PEAR-GLIO 
 
TITLE OF THE PROTOCOL:  Prospective Evaluation of AI R&D tool in adult Glioma and 
other primary brain tumours (PEAR-GLIO)  
 
IRAS Reference: 318531 
 
 
Sponsor:  
Ourotech Limited t/a Pear Bio 
 
Representative of the Sponsor: 
Duleek Ranatunga 
 
Finsgate, 5-7 Cranwood Street 
London, England 
EC1V 9EE 
 
Tel: 07716558079 
E-mail: duleek@pearbio.com 
 
 
 
Chief Investigator(s): 
Dr. Matt Williams 
 
Radiotherapy Dept 
Imperial College Healthcare NHS Trust 
Fulham Palace Rd 
W6 8RF 
 
EmaiL: matthew.williams@imperial.ac.uk 
Tel: 0203 3311 0733 
 
Principal Investigator(s): 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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STUDY CONTACTS 
 
Funder(s):  
 
Ourotech Limited (trading as Pear Bio), Finsgate, 5-7 Cranwood Street, London, England, EC1V 
9EE 
 
Statistical Advice: 
 
Pear Bio, Imperial College White City Campus, 84 Wood Lane, London, W12 0BZ.  
Contact: Dr. Francesco Iori, PhD 
E: francesco@pearbio.com  
 
Sponsor Laboratory: 
 
Pear Bio, Imperial College White City Campus, 84 Wood Lane, London, W12 0BZ.  
Contact: Dr.  Eleonora Peerani, PhD 
E: eleonora@pearbio.com  
Tel: (+39) 3319571756 
 
Trial Coordination: 
 
Pear Bio, Imperial College White City Campus, 84 Wood Lane, London, W12 0BZ.  
Contact: Ms. Elli Tham 
E: elli.tham@pearbio.com  
Tel: 07976094236 
 
Participating Sites: 
 
A list of participating sites can be obtained from the PEAR-GLIO Trial Coordinator 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  

 
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PROTOCOL SIGNATURE PAGE 
 
Signature of the Chief Investigator:  
 
The clinical study as detailed within this research protocol (Version 1.0, Dated 3 March 2023), 
and any subsequent amendments, will be conducted in accordance with the Research 
Governance Framework for Health & Social Care (2005), the World Medical Association 
Declaration of Helsinki (1996), Principles of ICH-GCP, and the current regulatory 
requirements, as detailed in the Medicines for Human Use (Clinical Trials) Regulations 2004 
(UK S.I. 2004/1031) and any subsequent amendments of the clinical trial regulations. 
 
Chief Investigator Name: 
Dr. Matthew Williams  
 
Signature and Date:  
______________________     __ __ / __ __ __ / __ __ __ __ 
 
 
Signature of Statistical Advisor:  
 
The clinical study as detailed within this research protocol (Version 1.0, Dated 3 March 2023), 
or any subsequent amendments, and will be conducted in accordance with the Research 
Governance Framework for Health & Social Care (2005), the World Medical Association 
Declaration of Helsinki (1996), Principles of ICH E6-GCP, ICH E9 -  Statistical principles for 
Clinical Trials, ICH E10 - Choice of Control Groups and the current regulatory requirements, 
as detailed in the Medicines for Human Use (Clinical Trials) Regulations 2004 (UK S.I. 
2004/1031) and any subsequent amendments of the clinical trial regulations. 
 
Name: Francesco Iori  
 
 
 
 
Signature and Date:  
______________________     __ __ / __ __ __ / __ __ __ __ 
 
 
Signature of the Principal Investigator:  
 
The clinical study as detailed within this research protocol (Version 1.0, Dated 3 March 2023), 
and any subsequent amendments, will be conducted in accordance with the Research 
Governance Framework for Health & Social Care (2005), the World Medical Association 
Declaration of Helsinki (1996), Principles of ICH-GCP, and the current regulatory 
requirements, as detailed in the Medicines for Human Use (Clinical Trials) Regulations 2004 
(UK S.I. 2004/1031) and any subsequent amendments of the clinical trial regulations. 
 
Name:  
 
 
 
Signature and Date:  
______________________     __ __ / __ __ __ / __ __ __ __ 
 
 
 
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TABLE OF CONTENTS 
STUDY CONTACTS………………………………………………………………………………….1 
PROTOCOL SIGNATURE PAGE…………………………………………………………………..2 
TABLE OF CONTENTS……………………………………………………………………………..3 
GLOSSARY OF TERMS AND ABBREVIATIONS……………………………………………….5 
STUDY SYNOPSIS…………………………………………………………………………………..7 
1. INTRODUCTION…………………………………………………………………………….9 
1.1 Trial outline 
 
1.2 Background and rationale 
 
1.2.1 
Current management of malignant gliomas  
1.2.2 
Treatment response in malignant gliomas 
1.2.3 
Predictive biomarkers of patient response 
1.3 Benefit/risk assessment 
 
2. STUDY AIMS AND OBJECTIVES……………………………………………………….12 
2.1 Primary objectives and endpoints 
2.2 Secondary objectives and endpoints  
2.3 Exploratory objectives and endpoints 
3. INVESTIGATIONAL PLAN………………………………………………………………..14 
3.1 Overall design  
3.2 Trial schema  
3.3 Data and tissue collected 
3.4 Laboratory schema   
3.5 Patient evaluability 
3.6 Replacement of patients 
3.7 Target accrual  
 
4. PATIENT SELECTION…………………………………………………………………….18 
4.1 Inclusion criteria 
 
4.2 Exclusion criteria 
 
5. STUDY PROCEDURES AND SCHEDULE OF ASSESSMENTS……………………19   
5.1 Patient identification  
5.2 Informed consent procedure  
5.3 Patient enrolment 
 
5.4 Schedule of assessments 
 
5.5 Procedures and measurements 
5.5.1 
Demographics and medical history  
5.5.2 
Height, weight and performance status 
5.5.3 
Concomitant medication 
5.5.4 
Haematology and clinical biochemistry 
 
5.5.5 
Treatment details 
5.5.6 
Tumour size evaluation 
 
5.5.7 
Adverse events 
 
5.5.8 
Surgical biopsy and blood collection 
5.5.9 
Exposure to Ionizing radiation 
 
5.6 Exploratory research 
5.6.1 
Chain of custody of biological samples 
5.7 Patient withdrawal 
 
6. PHARMACOVIGILANCE………………………………………………………………….25          

 
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6.1 Definition of an Adverse Event (AE)  
6.2 Recording of AEs 
 
6.3 Severity of AEs 
6.4 Causality of AEs 
 
6.5 Abnormal laboratory test results 
6.6 Definition of Serious Adverse Event (SAE) 
6.7 Reporting of SAEs 
6.7.1 
Non-reportable events 
6.8 Definition of an Adverse Reaction (AR) 
6.9 Definition of Serious Adverse Reaction (SAR) 
6.10 
Definition of Suspected Unexpected Serious Adverse Reaction 
(SUSAR) 
6.11 
Reporting of SUSARs 
6.12 
Annual reporting 
6.13 
Urgent safety measures 
 
7. STATISTICAL 
CONSIDERATIONS………………………………………………………28 
7.1 Sample size 
7.2 Statistical analysis 
 
7.2.1 
Primary efficacy analysis 
7.2.2 
Secondary efficacy analysis 
7.2.3 
Exploratory analyses 
7.3 Interim analysis and study termination 
7.4 End of study definition 
7.5 Handling of missing data 
 
8. DATA HANDLING AND RECORD KEEPING…………………………………………..31    
8.1 Confidentiality 
8.2 Study documents 
8.3 Data and sample acquisition  
8.4 Record retention and archiving 
8.5 Compliance 
 
9. STUDY MANAGEMENT…………………………………………………………………..33     
10. CLINICAL GOVERNANCE ISSUES……………………………………………………..33      
10.1 
Ethical considerations 
10.2 
Summary of monitoring plan 
10.3 
Audit and inspection 
10.4 
Reporting of serious breaches in GCP or the trial protocol  
11. STUDY FINANCES………………………………………………………………………34     
11.1 
Funding sources 
11.2 
Patient expenses/payments  
12. SPONSORSHIP AND INDEMNITY………………………………………………………34      
13. PUBLICATION POLICY…………………………………………………………………...35      
14. REFERENCES……………………………………………………………………………..35    
List of Tables and Figures in the Text 
Tables 
Table 1: Schedule of assessments  
Table 2: ECOG performance status  

 
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Figures 
Figure 1: Trial schema 
 
Figure 2: PEAR-GLIO laboratory workflow 
GLOSSARY OF TERMS AND ABBREVIATIONS 
 
AE 
Adverse Event 
ANC 
Absolute Neutrophil Count 
APR 
Annual Progress Report 
AST 
Aspartate aminotransferase 
CI 
Chief Investigator 
ECOG 
Eastern Cooperative Oncology Group 
eCRF 
Electronic Case Report Form 
EU 
European Union 
FBC 
Full Blood Count 
GCP 
Good Clinical Practice 
HTA 
Human Tissue Authority 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
IF 
Immunofluorescence  
ISF 
Investigator Site File 
NCI-CTCAE 
National Cancer Institute Common Toxicity Criteria For Adverse Events 
NRES 
National Research Ethics Service 
ORR 
Overall response rate 
OS 
Overall survival 
PD-(L)1 
Programmed death (ligand) 1 
PFS 
Progression free survival 
PI 
Principal Investigator 
PIS 
Patient Information Sheet 
REC 
Research Ethics Committee 
RNASeq 
RNA sequencing 

 
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SAE 
Serious adverse event 
SAR 
Serious adverse reaction 
SDV 
Source data verification 
SOP 
Standard Operating Procedures 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
TMB 
Tumour mutational burden 
TMF 
Trial Master File 
TMG 
Trial Management Group 
US 
Ultrasound 
WBC 
White blood cell count 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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STUDY SYNOPSIS 
 
Title 
Prospective Evaluation of AI R&D tool for Biology and Response in 
Adult Invasive Neuro-oncology (PEAR-GLIO) 
Main Objectives 
The study objectives are concerned with measurements and 
endpoints collected from laboratory testing on patient-derived 
primary brain tumours. 
 
The primary objective is to establish a functional dose of each FDA-
approved therapy for primary solid brain tumours in Pear Bio’s ex 
vivo platform, and to confirm these therapies demonstrate their 
intended mechanism of action (direct cell killing, cell killing by 
immune cell activation, etc.) This objective is quantified using intra-
patient and inter-patient statistics to evaluate differential ex vivo 
treatment efficacy for approved glioma therapies.  
 
Other objectives include measuring the correlation between multi-
omic biomarkers and ex vivo treatment response, and exploring 
responses to other potential therapies for brain tumours. 
 
Our Patient & Public Involvement & Engagement (PPIE) 
workstream will run in parallel, but will be open to a wider group of 
people, who may or may not be patients who participate in PEAR-
GLIO, and will be managed outside the study. 
Phase 
N/A 
Design 
This is a UK-based, observational study that aims to validate a 
diagnostic tool on its ability to test therapeutic sensitivity of brain 
tumours ex vivo. Patients diagnosed with brain tumours will 
undergo standard of care surgery, and have 40mL of whole blood 
collected. The blood and tissue material will be shipped fresh. The 
patient will go through routine post-surgical care and data 
collection. Any known treatments and events occurring post-
surgery, such as the administration of adjuvant therapy or 
recurrence, will be recorded and communicated to Pear Bio. The 
samples and data sent to Pear Bio will be used to evaluate the 
study’s objectives. 

 
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Sample Size 
50 
Inclusion Criteria 
1. Patient diagnosed with operable brain cancer, thought likely 
to be primary solid brain tumour on imaging (grade 2 - 3 
meningioma; grade 1 - 4 tumours otherwise) or with 
histologically proven primary malignant solid brain tumour; 
2. Able to give written informed consent prior to admission to 
this study; 
3. Female or male aged ≥18 years; 
4. Patient consents to the use of their surgical sample and 
40mL of whole blood for research purposes; 
5. Surgical sample and yields ≥0.4g for the study; 
6. Patient consents to providing histopathology data (e.g., 
confirmation of histological subtype as oligodendroglioma) 
and other pseudonymised health information including 
imaging, treatment and outcome data. 
 
Exclusion Criteria 
1. Inoperable or biopsy only; 
2. Suspected lymphoma or myeloma, or grade 1 meningioma 
3. Preoperative haemoglobin levels below 120g/L; 
4. Patients who have already received chemotherapy, 
targeted therapy, immunotherapy, or radiotherapy less than 
30 days before date of surgery, unless as part of a clinical 
trial (requires per-patient sponsor approval). 
5. Recurrence of cancer originating from a site other than the 
brain; 
6. Any other disease, metabolic dysfunction, physical 
examination finding, or clinical laboratory finding that, in the 
investigator’s opinion, gives reasonable suspicion of a 
disease or condition that may affect the interpretation of the 
results, render the patient at high risk from treatment 
complications or interferes with obtaining informed consent. 
 

 
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1: INTRODUCTION 
 
1.1 Trial outline 
Pear Bio have developed a 3D micro-tumour and computer vision platform to culture patient-
derived tumour samples and predict sensitivity to various therapeutic agents. This study is 
intended to validate the mechanism of action of FDA-approved primary brain tumour therapies, 
such as Temozolomide, Lomustine and Bevacizumab, as well as therapies being assessed in 
clinical trials, on fresh brain tumours and autologous immune cells isolated from whole blood. 
This is a proof of concept study to confirm that therapies used in primary brain tumours can 
be tested in Pear Bio’s platform. Follow-on studies will be used to determine the sensitivity 
and specificity of Pear Bio’s test in predicting patient response (OS and PFS). This study will 
acquire fresh patient tumour samples and 40 mL of whole blood per patient. Samples will be 
shipped fresh on the same day of collection to Pear Bio’s lab. Histopathology reports will be 
provided to Pear Bio shortly after the sample shipment in order to confirm the tumour subtype 
(although samples will not be excluded from analysis based on tumour subtype). Other 
available patient data, including imaging and imaging reports, will also be provided in a 
pseudonymised form. Patients who have had previous tissue sampling (i.e. those undergoing 
treatment for relapsed disease) will also provide information on previous histology and 
treatment. 
Pear Bio will isolate cells from the tumour samples and whole blood, co-culture them into 3D 
micro-tumours, and run live microscopy-based assays to determine ex vivo tumour response 
to treatments. Image data will be analysed using a proprietary computer vision pipeline to 
measure ex vivo tumour response metrics, such as tumour cell death, tumour cell migration 
and immune cell infiltration. Molecular biology assays will be conducted to determine changes 
in DNA features (tumour mutational burden and mis-match repair), gene expression (PCR or 
RNASeq) and protein distribution (immunofluorescence or spatial proteomics assays) for 
biomarkers due to ex vivo treatment exposure. 
Various Pear Bio laboratory assays will be compared to validate cell culture models, computer 
vision algorithms and drug mechanisms of action. Any available patient data will also be used 
to determine whether Pear Bio’s assays can predict patient outcomes, such as recurrence or 
eligibility for future targeted therapies. However, due to the variety of patients recruited, the 
low total sample size, and the lack of evaluable patients receiving non-surgical treatment, no 
formal statistical analysis will be conducted on patient outcomes. 
At the end of this study, Pear Bio will determine whether the platform has potential for  patient 
stratification in primary brain tumours. Future studies will aim to demonstrate the 
sensitivity/specificity of Pear Bio’s platform for an eventual intended use in guiding treatment 
decision making for patients with primary brain tumours in order to increase their response 
rates.  

 
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Our Patient & Public Engagement & Involvement (PPIE) workstream will hold three meetings 
and use online questionnaires and consultations to help understand patient, carer, 
professional and public views on the Pear Bio platform and its use. 
1.2 Background and rationale 
 
Brain tumours are a  large unmet need in the UK and worldwide. They are the leading cause 
of cancer death in the under-40’s, with 12,500 new patients diagnosed annually in the UK and 
88% dying within 5 years (1) . The US faces similar problems, with 79,000 new cases and 
nearly 14,000 deaths annually (2). Of the malignant brain tumours, glial tumours are the most 
common, making up 70% of all adult malignant brain tumour diagnoses.  
 
1.2.1 Current management of malignant glioma 
 
Malignant glioma includes grade 2-4 glioma, and forms the majority of primary brain tumours 
in adults. Glioma therapy depends on grade and subtype but broadly includes a combination 
of surgical resection, radiotherapy and chemotherapy. 
 
Optimal treatment of glioblastoma (grade 4 glioma) consists of surgery, chemo-radiotherapy 
and chemotherapy with Temozolomide (3). For grade 3 astrocytoma, surgery, radiotherapy 
and adjuvant temozolomide are standard of care (4), and for oligodendroglioma, surgery, 
radiotherapy and PCV-based chemotherapy are standard of care. At relapse, therapy is 
typically further chemotherapy, either with the same agents, or previous unused 
chemotherapy. 3rd-line chemotherapy is poorly defined, but might include carboplatin or 
irinotecan, and may include bevacizumab. 
 
Combined TMZ & Lomustine may be beneficial in MGMT methylated GBM, but apart from 
MGMT methylation, IDH mutation and 1p/19q co-deletion - which predict response to 
chemotherapy in general - there are no biomarkers to predict response to specific therapeutic 
agents. 
 
1.2.2 Treatment response in malignant primary brain tumours 
 
Primary brain tumours have a 5-year survival rate of only 12%, and GBM has a median survival 
of 15 months with aggressive multimodal treatment (5). However, there is some evidence to 
support use of combined chemotherapy in newly diagnosed patients with GBM, uncertainty 
about chemotherapy regimen choice in grade 2 and 3 gliomas, and uncertainty about therapy 
choice at relapse across all gliomas. 
 
1.2.2 Other primary brain tumours 
 
Outside of the gliomas, the commonest primary brain tumour is a grade 1 meningioma. We 
are not including these patients in the study, as the tumours have very good outcomes, and 
are unlikely to be possible to grow in our system. However, there are a range of other primary 
brain tumours, all of which are much rarer (e.g. ependymoma, grade 1 gliomas) which would 
all be eligible for inclusion in the study. 
 

 
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1.2.3 Predictive biomarkers of patient response 
 
The only good predictive biomarkers for glioma are IDH mutation status, 1p/19q codeletion 
and MGMT methylation. However, these all generally predict for chemotherapy 
responsiveness, rather than which therapy is best. 
 
Pear Bio’s test provides a potential avenue to test single agents and combination therapies 
prior to treatment selection to guide decision-making.  
 
 
1.3 Benefit/risk assessment 
 
This is an observational study with patients receiving standard of care surgery. Samples 
collected from surgery will be used in Pear Bio’s test, but they will not guide any treatment 
decisions. As such, there are no benefits to the patients taking part in this study.  
 
This study will be used to conduct analytical validation of the patient stratification tool. The 
study data will be used to inform the design of future trials, which will be aimed at increasing 
patient response rates by using the test before systemic therapy starts to decide on the optimal 
treatment regimen(s) to use for an individual with glioma. 
 
As surgery and blood collection are within routine care for resectable brain cancer, study-
specific procedures do not pose significant risks to patients. A source of potential risk comes 
from collecting 40mL of matched whole blood per patient, which may go beyond routine care 
to satisfy study requirements. Possible risks associated with extra blood collection are a 
feeling of lightheadedness, dizziness and local bruising. However, this will be mitigated as 
patients with preoperative haemoglobin levels below 120g/L will not be recruited. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
2: STUDY AIMS AND OBJECTIVES 
 
 
2.1 Primary objectives and endpoints 
 

 
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Primary objective 
Endpoints 
The primary objective is to 
establish a functional dose of 
each FDA-approved therapy for 
glioma in Pear Bio’s ex vivo 
platform, and to confirm these 
therapies 
demonstrate 
their 
intended mechanism of action 
(direct cell killing, cell killing by 
immune cell activation, etc.) 
As the primary objective is laboratory-based, a patient 
outcome endpoint is not necessary. Instead, success will 
be determined by: 
 
1. Observing 
differentiated 
ex 
vivo 
treatment 
response across the therapies/combos tested on 
each patient’s tumour sample (intra-patient 
sample comparison) 
2. Observing 
differentiated 
ex 
vivo 
treatment 
response levels between the cohort of samples 
collected 
from 
patients 
on 
a 
per 
therapeutic/combo basis (inter-patient sample 
comparison) 
 
2.2 Secondary objectives and endpoints 
 
Secondary objectives 
Endpoints 
Assess the accuracy of Pear 
Bio’s assay at correlating to 
patient progression free 
survival (PFS)   
Tumour response will be measured as clinically indicated 
(typically every 3 months), and at all subsequent 
timepoints until disease progression, as per standard of 
care.  
 
Disease progression is evaluated by the patient’s 
radiologist and is defined by RANO guidelines. 
 
Kaplan–Meier curves will be generated on the patient 
population, and where feasible, based on their line of 
treatment and for each therapeutic option (if n is 
sufficient). These curves will be compared to reported 
data to determine how representative the patient 
population is of past trials and clinical practice.  
 
Computer vision biomarkers will be categorised into 
low/high groups to determine their correlation with PFS. 
This analysis will not demonstrate the patient benefit of 
using Pear Bio’s tool, but it will generate hypotheses for 
interventional trials designed to demonstrate patient 
benefit.  
 
 
 
 
 
 
2.3 Exploratory objectives and endpoints 
 
Tertiary objectives 
Endpoints 

 
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Determine 
the 
rate 
of 
successfully 
established 
cultures from surgical samples 
The percentage of cultures in which ≥70% of viable 
tumour cells encapsulated post-isolation on day 0 are still 
alive on day 3 in the control cultures (no treatment) 
compared to the number of tumour samples (≥0.4g) 
successfully accepted at Pear Bio’s lab.   
Determine 
the 
rate 
of 
successfully cultured immune 
cells 
The percentage of cultures in which ≥70% of viable 
immune cells plated post-extraction on day 0 are still alive 
on day 3 in the control cultures (no treatment) compared 
to the number of blood samples (≥40mL) successfully 
accepted at Pear Bio’s lab.   
Find correlations between ex 
vivo tumour culture or multi-
omic biomarkers and real-world 
patient outcomes 
For any cases where patient outcomes are available or 
become available prospectively, such as recurrence post-
surgery, potential predictive or prognostic biomarkers will 
be identified. As this data will be sparse, the analysis will 
only be used to generate hypotheses for future trials, if the 
data allows.  
Assess Pear Bio’s assay ability 
to categorise patients for 
below average or above 
average overall survival (OS)  
Patient data is collected up to death and their time from 
diagnosis to death is recorded to determine the overall 
survival (OS) time.  
 
This analysis will use median OS within the study cohort 
to differentiate patients as generally responsive or 
resistant to treatment. The analysis will then explore 
indicators/biomarkers in Pear Bio’s test that can identify 
patients as generally responsive or resistant to 
treatment, agnostic of the therapeutic choice.  
Assess the correlation of 
omics biomarkers to patient 
PFS, ORR and/or OS 
Omics readouts taken of patient tumour samples at Pear 
Bio’s laboratory will be used to determine whether any 
biomarkers can correlate to therapeutic response. 
 
Omics methods include: 
1. Immunofluorescence (IF) 
2. RNASeq  
3. Tumour mutational burden (TMB) and 
microsatellite instability (MSI) 
 
Prediction methods include: 
1. Correlating baseline expression of biomarkers in 
the tumour sample to real-world patient outcomes 
2. Comparing the change in biomarker expression 
after ex vivo treatment testing to real-world 
patient outcomes 
 
 
 
3: INVESTIGATIONAL PLAN 
 
 
3.1 Overall design 
 

 
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This is a UK-based, observational study that aims to validate a diagnostic tool on its ability to 
test therapeutic sensitivity of primary solid brain tumours ex vivo. Patients diagnosed with a 
brain tumour on imaging, or who have had previous treatment for a brain tumour and are now 
undergoing further surgery as clinically indicated, will undergo standard of care surgery, and 
have 40mL of whole blood collected. The material available from the surgical sample for this 
study will be shipped fresh alongside whole blood to Pear Bio. The patient will go through 
routine post-surgical care and data collection. Any previous or subsequent treatments and 
events occurring post-surgery, such as the administration of adjuvant therapy or recurrence, 
will be recorded and communicated to Pear Bio. The samples and data sent to Pear Bio will 
be used to evaluate the study’s objectives.  
 
3.2 Trial schema 
 
 
Figure 1: Trial schema 
 
 
3.3 Data and tissue collected 
 
 
Biospecimens 
Data 
Required 
1. At least 0.4g of 
fresh, unfixed, 
tumour tissue taken 
from surgery 
2. 4 x 10mL EDTA 
vials of matched 
whole blood 
1. Demographic data 
(pseudonymised) 
2. Redacted pathology reports, 
including immunohistochemistry 
tests, molecular pathology and 
NGS (IDH and MGMT in 
particular) and further genomic 
data where available, including 
methylation profiling 

 
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3. Radiology images and reports 
4. Concomitant medications 
 
Collected if available 
N/A 
1. Blood and liver/kidney function 
tests at baseline 
2. Follow-up outcome data, 
including recurrences 
3. Follow-up treatment data, 
including adjuvant therapy 
regimens 
 
3.4 Laboratory setup 
Fresh tissue resections that arrive at Pear Bio’s lab will undergo processing, cell culture and 
various drug dosing and omics assays (depending on extracted cell numbers). Tumour 
samples will be processed using a cell isolation kit to retrieve a viable single-cell suspension. 
A minimum of 100,000 viable cells will be used for staining with live and dead cell-tracking 
dyes. In parallel, blood vials will be processed for PBMCs and further effector cell extraction 
(flow cytometry, Dynabeads, etc). The remaining cells will be used for various omics assays 
including looking at DNA alterations (tumour mutational burden), gene expression (PCR or 
RNASeq) and protein/receptor distribution (immunofluorescence or spatial proteomics assays 
for biomarkers such as PD-(L)1).  Wherever sample size allows it, tissue chunks are cut out 
of the main tumour bulk prior to cell isolation for spatial analyses (GeoMX Nanostring). 
The stained cells will be cultured in a biomimetic hydrogel within Pear Bio’s 3D micro-tumour 
platform to provide a physiological environment for drug dosing experiments.  Therapies will 
be administered to each 3D micro-tumour over multiple days  (either as monotherapy or 
combination therapies as outlined below) . 
 
Standard set of therapies tested 
3D cell culture 1 
Control 
3D cell culture 2 
Temozolomide 
3D cell culture 3  
Lomustine 
3D cell culture 4 
Temozolomide + Lomustine 

 
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3D cell culture 5 
Procarbazine + Lomustine 
3D cell culture 6 
Carboplatin 
If enough cells are available, we will test additional therapies including Etoposide, 
Regorafenib, Pembrolizumab, Irinotecan, Avastin, Olaparib, Nariparib, Lapatinib,  
Entrectinib, Osimertinib and others, either alone or in combination. 
 
In parallel, PBMCs will be extracted from whole blood, characterised via flow cytometry and 
sorted via fluorescence-activated cell sorting (FACS) or magnetic beads selection. Cells of 
interest (e.g. CD8+ T cells) will be used for culture in Pear Bio's 3D system jointly with cells 
isolated from the matched tumour sample. Tumour-isolated cells will be co-cultured with 
immune cells. Cultures receiving immunotherapies may be tested for tumour mutational 
burden. 
Confocal microscopy will be conducted daily to collect 3D image data of the cells and track 
their position and behaviour over time. At the end of the assay, the 3D cell cultures will be 
fixed or snap-frozen for further 3D immunofluorescence analyses or used for embedding, 
sectioning and assessment of spatial proteomics. For targeted therapies, RNAseq, IF and 
proteomics data will be integrated to confirm drug MoA and identify other potential therapeutic 
targets. Concurrently, 3D image data will be processed through a computer vision pipeline to 
measure functional metrics of the ex vivo 3D cell cultures, including cell viability, tumour 
culture width and cell migration, both at a bulk tumour level and at a single-cell resolution. A 
patient report is generated outlining an individual patient sample’s response to each therapy 
tested.  
Potential additional analyses: 
● 
DNA analyses: 
o 
Tumour mutational burden (TMB) 
o 
Microsatellite instability (MSI) 
o 
Fluorescent in-situ hybridisation (FISH) 
o 
Genome-wide methylation patterns 
o 
Whole genome sequencing 
o 
Whole exome sequencing 
 
● 
RNA analyses: 
o 
RNASeq 
o 
Microarrays 
o 
Quantitative reverse transcription PCR (RT-qPCR) 
o 
Spatial analyses (Nanostring, ResolveBiosciences) 
● 
Protein analyses: 
o 
Flow cytometry (FC) 
o 
Immunofluorescence (IF) 
o 
Analysis of secreted factors 

 
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Figure 2: PEAR-GLIO laboratory workflow. (1) Patient gives a pre-op blood sample (2) Patient 
undergoes surgery as part of standard of care (3) Viable cells are isolated from the tumour and immune 
cells are isolated from blood samples (4) Tumour-dissociated cells  and immune cells are stained with 
live-cell and dead-cell fluorescent dyes. (5) The cells are cultured simultaneously in multiple Pear Bio 
hydrogels contained in a microfluidics device. (6) Standard-of-care chemotherapy and targeted 
treatments are dosed within the device whilst (7) daily imaging allows for live tracking of cell viability 
and migration. (8) Wherever additional cells are available, omics assays (e.g. RNA sequencing and 
immunofluorescence) will be run in parallel to check expression of common biomarkers and validate 
drug mode of action. (9) Computer vision (CV) is implemented to detect changes in cell morphology, 
viability, and position (amongst other parameters) over time in order to make an informed prediction of 
differential treatment efficacies. 
 
3.5 Patient evaluability 
 
 
In order to be considered evaluable, patients must meet the eligibility criteria, consent to the 
use of their surgical sample and blood for the study, and yield a research sample with  
sufficient weight and tumour cell content resulting in the establishment of a successful cell 
culture in the Pear Bio laboratory. 
 
3.6 Replacement of patients 
 
Patients who do not meet the evaluability criteria set out in section 3.5 will be replaced. 
 
3.7 Target accrual 
 
 
A maximum of 50 evaluable patients will be recruited in this study. On recruitment of the first 
10 patients, the Trial Management Group (TMG) will meet to assess whether monthly 
recruitment targets are met, and to confirm sample quality and successful culture rates upon 
processing at the Pear Bio laboratory.  
 
The TMG will use the results to determine whether to increase accrual up to a maximum of 50 
patients. 
 
 
4: PATIENT SELECTION 
 
4.1 Inclusion criteria 
 

 
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1. Patient diagnosed with operable brain tumour, thought likely to be primary solid brain 
tumour on imaging (grade 2 - 3 meningioma or grade 1 - 4 for other tumours) or with 
histologically proven primary malignant solid brain tumour and due to undergo surgery 
as clinically indicated 
2. Able to give written informed consent prior to admission to this study; 
3. Female or male aged ≥18 years; 
4. Patient consents to the use of their surgical sample and 40mL of whole blood for 
research purposes; 
5. Surgical sample yields ≥0.4g for the study; 
6. Patient consents to providing histopathology data (e.g., confirmation of histological 
subtype as oligodendroglioma) and other pseudonymised health information including 
imaging, treatment and outcome data. 
 
4.2 Exclusion criteria 
 
1. Inoperable or biopsy only 
2. Suspected lymphoma or myeloma, or grade 1 meningioma; 
3. Preoperative haemoglobin levels below 120g/L; 
4. Patients who have already received chemotherapy, targeted therapy, immunotherapy, 
or radiotherapy less than 30 days before date of surgery, unless delivered as part of a 
clinical trial (must be discussed with sponsor on a per-patient basis) 
5. Recurrence of cancer originating from a site other than the brain; 
6. Any other disease, metabolic dysfunction, physical examination finding, or clinical 
laboratory finding that, in the investigator’s opinion, gives reasonable suspicion of a 
disease or condition that may affect the interpretation of the results, render the patient 
at high risk from treatment complications or interferes with obtaining informed consent. 
 
 
 
 
 
 
 
 
 
 
 
5: STUDY PROCEDURES AND SCHEDULE OF ASSESSMENTS 
 
5.1 Patient identification 
 
Patients will be identified in multi-disciplinary team meetings or in outpatient clinics by their 
clinical care team.  
 
Only individuals with legitimate access to medical records (such as the patient's clinical care 
team) will be accessing these records for the purpose of identifying participants.  
 

 
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5.2 Informed consent procedure 
 
 
It is the responsibility of the Investigator, or a medically trained person delegated by the 
Investigator to obtain written informed consent from each subject prior to participation in this 
study, following adequate explanation of the aims, methods, anticipated benefits and potential 
hazards of taking part in the study. Ample time must be given for consideration by the patient 
before taking part. Attempts will be made to arrange for an official hospital translator for any 
participant who is not competent or comfortable with communication in English. The translator 
will be asked to read through the Patient Information Sheet (PIS) and Consent Form and to 
translate each section for the participant. Written informed consent will only be obtained from 
those who the Investigator feels assured have understood the implications of participation in 
the study. Patients with mental capacity issues will not be included in this study.  The PI must 
document in the patient’s notes when the PIS was given to the patient and when informed 
consent was obtained. 
 
If new safety information becomes available, the CI, in conjunction with the TMG, will review 
the study, update the PIS accordingly and resubmit for relevant approvals. The CI will review 
the new safety information and assess whether an urgent TMG meeting should be convened 
or whether this information can be reviewed at the next scheduled meeting. All patients, 
including those already undergoing scans, should be informed of the new information, given 
a copy of the revised PIS, and asked to give their consent to continue in the study. Patients 
will not be re-consented following amendments that do not affect safety or the number of 
assessments/visits required.  
 
5.3 Patient enrolment 
 
Principal Investigator(s) (PIs) at each recruiting site must keep a record of all patients 
screened for entry into this study, including those deemed ineligible after screening. Copies of 
the screening logs should be filed in the Investigator Site File (ISF). For each patient, the 
primary reason for exclusion should be recorded. Diagnostic data obtained as part of the 
patient’s standard care can be used to determine eligibility provided they fall within the protocol 
defined timelines. Written informed consent must be obtained prior to the patient undergoing 
any study-specific procedures.   
 
After ensuring that a patient has consented to participate in the study, a registration electronic 
case report form (eCRF) must be completed. Patients will then undergo screening to confirm 
study eligibility. Once it has been confirmed that a patient meets all eligibility criteria, the study 
site will submit the patient’s eligibility information to the coordinating centre. The clinical site 
will assign patients with a unique study ID for use in all correspondences (the Sponsor will 
provide a sequence of codes to assign). To ensure patient confidentiality, patients will only be 
identified using their assigned study ID on eCRFs, other study specific forms and all 
communications to the Sponsor. It is the PI’s responsibility to maintain a confidential record of 
the identity (i.e., full name, date of birth and hospital number) for the patients enrolled in this 
study and their assigned study ID. At the end of the study, this record should be archived along 
with the ISF.   
 

 
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Full details of how to enrol a patient via the PEAR-GLIO eCRF can be found in the eCRF 
completion guidance document. 
 
5.4 Schedule of assessments 
 
Patients may not need to make any additional site visits as the samples collected for the study 
can be taken from standard of care procedures. Due to logistical reasons, it may be difficult 
for the recruiting site to carry out all screening assessments in one day. Patients will be fully 
informed about the number of visits required to confirm eligibility in the trial. Subsequent visits 
will be as per standard of care at the local institution.  For a summary of assessments see 
Table 1. 
 
 
Baseline 
(Before 
surgery) 
At or shortly 
after surgery 
 
Standard of care 
follow-ups 
 
 
If cancer recurs 
Informed 
consent 
and 
eligibility 
checks 
B 
 
 
 
Demographics 
and 
medical 
history 
A 
 
 
 
Height, 
weight 
ECOG 
A 
 
 
 
Concomitant 
medication 
A 
 
 
 
Results 
from 
standard of care 
haematology, 
biochemistry 
assessments 
A 
 
 
 
Standard of care 
imaging 
A 
 
 
 
Tumour 
size 
evaluation 
A 
 
 
 
Standard of care 
surgery 
 
 
A 
 
 
Histopathology1 
 
A 
 
A 
Collection 
of 
blood 
for 
the 
study2 
 
B (40mL whole 
blood) 
 
 
Collection 
of 
tissue and data 
for the study2 
 
B 
 
 

 
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Transfer of data 
for the study2 
 
B 
B 
B 
Adverse Events 
by CTCAE v5.03 
 
B 
 
 
Treatment 
and 
follow-up 
as 
clinically 
indicated 
 
 
A 
 
Follow-up 
assessments for 
recurrence4 
 
 
A 
 
Further therapy5 
 
 
 
A 
A = Standard of care assessment 
B = Study-specific assessment or data collection 
Table 1: Schedule of assessments 
 
Table notes: 
1. A redacted histopathology report will be sent to Pear Bio when it is ready (i.e., it does 
not have to be sent alongside the tissue, blood and other available data from the day 
of the surgery). 
2. Available data is transferred alongside the tumour sample within 24 hours of surgery 
to Pear Bio’s laboratory. Any remaining data will be transferred as soon as feasible, 
and we will transfer any further imaging at a single timepoint at 12 months after surgery  
3. Relating only to research procedures (e.g., taking a volume of blood that is beyond 
standard care for a given patient). This can be conducted by telephone – physical 
examination to be done only if clinically indicated. 
4. Collected only if available for consenting patients within the study period. 
5. Collected only if available for consenting patients within the study period. 
 
 
 
5.5 Procedures and measurements 
 
5.5.1 Demographics and medical history 
 
Demographic data collected will include age, sex and race/ethnicity. Details of medical history 
obtained as part of standard of care will be collected, including details of any relevant medical 
conditions occurring prior to consent.  
 
Details will also be collected on the patient’s cancer diagnosis, including site, date of 
diagnosis, and tumour size. 
 
5.5.2 Height, weight and performance status 
 
Baseline height (cm) and weight (kg) will be collected from the medical records.  

 
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Performance status data will be collected at baseline using the ECOG performance score 
according to Table 2. This information will be recorded in the e-CRF. 
 
Grade 
ECOG 
0 
Fully active, able to carry on all pre-disease performance without restriction 
1 
Restricted in physically strenuous activity but ambulatory and able to carry out 
work of a light or sedentary nature, e.g., light house work, office work 
2 
Ambulatory and capable of all self-care but unable to carry out any work activities. 
Up and about more than 50% of waking hours 
3 
Capable of only limited self-care, confined to bed or chair more than 50% of waking 
hours 
4 
Completely disabled. Cannot carry on any self-care. Totally confined to bed or 
chair 
5 
Dead 
Table 2: ECOG performance status 
 
5.5.3 Concomitant medication 
 
All medications (including prescription medications and over the counter preparations) taken 
by the patient during the screening period will be documented as concomitant medications. 
The following details will be collected at baseline: drug name, reason for treatment, dose/units, 
route of administration, frequency. 
 
5.5.4 Haematology and clinical biochemistry 
 
 
The results of any standard of care haematology and clinical biochemistry tests will be 
collected at baseline. The date and result for each test must be recorded in the appropriate 
eCRF.  
 
5.5.5 Treatment details 
 
Patients will receive surgery as per standard of care. Consenting patients will have a portion 
of their tumour and blood used for this study. 
 
For patients who have adjuvant therapy within the study period, the following details will be 
collected at each cycle: drug name, start date and end date, dose/units, dose 
reductions/interruptions, reasons for any treatment changes/interruptions/dose reductions, or 
details of radiotherapy dose, fractionation, start and end dates, and for surgery, dates and 
times of surgery and post-operative stay. 
 
5.5.6 Tumour size evaluation 

 
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Measurements will be made by contrast-enhanced MRI. Imaging reports and pseudonymised 
images will be included as study data collected and transferred to Pear Bio, provided the 
imaging was done within the period of 3 months prior to surgery or during the follow-up period. 
Sites will transfer initial imaging shortly after surgery, and again at a single timepoint 12 months 
after surgery. 
 
5.5.7 Adverse events 
 
Adverse events will be restricted to those resulting from study-related procedures (e.g., a 
volume of blood collected beyond standard care to satisfy study requirements). The following 
details will be collected: AE term, date of onset, date of resolution, CTCAE grade (maximum 
intensity), seriousness, investigator causality rating against research procedures (yes or no), 
action taken with regard to the research procedures and outcome. 
 
5.5.8 Surgical biopsy and blood collection 
 
40mL of whole blood will be collected in 4x 10mL EDTA tubes on the day of surgery.  
 
At the time of surgery, a portion of the tumour resected from surgery will be provided to Pear 
Bio (≥0.4g). Samples must be placed in tissue transport medium to be supplied by Pear Bio. 
 
Each sample should be stored at 4°C before being transported by express courier to Pear Bio 
so that samples arrive within 24 hours of collection; blood and tumour samples may be sent 
separately because they may be collected up to 4 days apart.  
 
5.5.9 Exposure to Ionizing radiation 
 
There are no expected research exposures to ionising radiation. Although patients may 
undergo imaging with ionising radiation, and many may undergo radiotherapy, all of these will 
be delivered as part of routine care, and not as part of the trial. In addition, disease 
assessments are routinely carried out using MRI, which is a non-ionising modality. 
 
 
 
5.6 Exploratory research 
 
All patients will be consented for the collection and use of their tumour tissue and blood 
samples. All samples will be link-anonymised and only identified by the study ID and unique 
sample number allocated by the clinical site (Sponsor to provide sequence of codes to assign). 
These results may be reported separately from the clinical study report. 
 
5.6.1 Chain of custody of biological samples 
 
In all cases, patients will be consented for the collection and use of their biological samples 
and a full chain of custody will be maintained for all samples throughout their lifecycle. The 
Investigator at each site is responsible for maintaining a record of full traceability of biological 

 
PEAR-GLIO Protocol v1.0 
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samples collected from patients while these are in storage at the site; either until shipment or 
disposal. Any sample receiver (e.g., sub-contracted service provider) will keep full traceability 
of samples from receipt of arrival to further shipment or disposal (as appropriate). 
 
In the event that a patient withdraws their consent from the study, all samples and data 
collected up to that date will be used in the study, but no further data will be collected. As the 
Sponsor, Ourotech Limited (trading as Pear Bio) will maintain oversight of the entire lifecycle 
through internal procedures and monitoring of the study site(s). The Sponsor will be the 
custodian of the samples. Samples will be transferred from the participating site to Ourotech 
Limited (trading as Pear Bio). At the end of the study, unused samples (or portions of samples) 
will be retained for future research while all used samples (or portions of samples) will be 
disposed of in accordance with the Human Tissue Act 2004.  
 
5.7 Patient withdrawal 
 
Patients may voluntarily withdraw from the study at any time. Patients will also be withdrawn 
from the study if their research sample has insufficient weight (<0.4g) or tumour content (<100 
000 viable cells), or the sample fails to establish a culture in the laboratory. 
 
5.8 PPIE workstream 
 
The PPIE workstream will run separately from the trial: patients enrolled in the trial will be 
invited to participate in the PPIE work, but there will not be automatic overlap (i.e. patients will 
be invited to PPIE, but enrollment in the trial is independent of PPIE engagement, and PPIE 
engagement does not count towards trial enrollment).  
 
We will engage with brain tumour patients and carers, as well as professionals and the wider 
public by working with the local neuro-oncology PPI team. We will work with the local clinical 
team to identify participants, and participation will be by invitation and voluntary participation. 
All data collected will be anonymised and summarised for thematic analysis and discussion, 
and to inform the development of the next round of clinical trials. 
 
 
 
 
6: PHARMACOVIGILANCE 
 
6.1 Definition of an Adverse Event (AE) 
 
An AE is any untoward medical occurrence (including deterioration of a pre-existing medical 
condition) in a subject who is administered any research procedure, which does not 
necessarily have a causal relationship with this procedure. An AE can therefore be any 
unfavourable and unintended sign (including an abnormal laboratory finding), symptom or 
disease temporarily associated with a research procedure, whether or not considered related 
to the procedure.  
 
6.2 Recording of AEs 

 
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AEs will be collected throughout the study, from informed consent through post-surgical care. 
They will be followed up according to local practice until the event has stabilised or resolved. 
Any unresolved AEs at the patient’s last visit should be followed up for as long as medically 
indicated, but without further recording in the eCRF. The following details will be collected in 
the eCRF for each AE: AE term, date of onset, date of resolution, NCI-CTCAE grade maximum 
intensity, seriousness, investigator causality rating against research procedures, action taken 
with regards to research procedures and outcome.  
 
6.3 Severity of AEs 
 
Severity is a measure of intensity whereas seriousness is defined by the criteria in section 6.6. 
Severity will be assessed using the grading scales found in the National Cancer Institute 
CTCAE version v5.0 (27Nov2017) for all AEs with an assigned NCI-CTCAE term. For those 
events without assigned NCI-CTCAE grades, the recommendation on page 1 of the NCI-
CTCAE that converts mild, moderate and severe into NCI-CTCAE grades should be used. A 
copy of the NCI-CTCAE version 5.0 can be downloaded from the Cancer Therapy Evaluation 
Program website (http://ctep.cancer.gov). 
 
6.4 Causality of AEs 
 
The Investigator will assess causal relationships between research procedures and each AE. 
 
6.5 Abnormal laboratory test results 
 
Not applicable. Haematological and biochemical parameters will not be assessed throughout 
the study.  
 
6.6 Definition of Serious Adverse Event (SAE) 
 
An SAE is an AE occurring during any part of the study that meets one or more of the following 
criteria: 
● Is fatal – results in death  
○ NOTE: death is an outcome, not an event 
● Is life-threatening  
○ NOTE: The term ‘life threatening’ in the definition of ‘serious’ refers to an event 
in which the patient was at risk of death at the time of the event; it does not 
refer to an event which hypothetically might have caused death if it were more 
serious, 
● Requires inpatient hospitalisation or prolongation of existing hospitalisation 
○ NOTE: “Hospitalisation” means any unexpected admission to a hospital. It does 
not usually apply to scheduled admissions that were planned before study 
inclusion or visits to casualty (without admission). Elective admissions for 
surgery are also excluded. 
● Results in persistent or significant disability/incapacity 
● Is a congenital anomaly/birth defect 
● Other important medical events  

 
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○ NOTE: Medical judgement should be exercised in deciding whether an adverse 
event/reaction 
is 
serious 
in 
other 
situations. 
Important 
adverse 
events/reactions that are not immediately life-threatening, or do not result in 
death or hospitalisation but may jeopardise a subject, or may require 
intervention to prevent one of the other outcomes listed in the definition above 
should also be considered serious. 
 
6.7 Reporting of SAEs 
 
Rapid reporting of all SAEs occurring from consent until 3 days after study sample collection 
must be performed as detailed in the “SAE reporting instructions” within 24 hours of the PI or 
designee becoming aware of the event. If the investigator becomes aware of safety 
information that appears to be related to a research procedure involving a subject who 
participated in the study, even after an individual subject has completed the study, this should 
also be reported to the Sponsor. All SAEs should be reported to Sponsor using the SAE form 
and will be reviewed by the CI or designated representative to confirm relatedness and 
expectedness. Following documented assessment by a delegated investigator, the completed 
SAE form will be forwarded to the Sponsor by the clinical site within the pre-specified timelines. 
 
All SAEs must be reported to the Sponsor using the PEAR-GLIO SAE form via email and 
within 24 hours of the site becoming aware of the event. 
 
Please note all events should also be recorded in the relevant sections of the case report 
forms and patient medical records. 
 
6.7.1 Non-reportable events 
 
Due to the nature of the disease in this study, the following situations that fulfil the definition 
of an SAE are excluded from recording/reporting on an SAE form. However, they should be 
recorded on the eCRF and in the medical records. 
 
● Elective hospitalisation and surgery for treatment of cancer or its complications. 
● Prolonged hospitalisation for post-surgical complications or post anti-cancer treatment 
complications. 
● Elective hospitalisation to make treatment or procedures easier. 
● Elective hospitalisation for pre-existing conditions that have not been exacerbated by 
trial treatment. 
 
6.8 Definition of an Adverse Reaction (AR) 
 
An AR is any unfavourable and unintended sign (including an abnormal laboratory finding), 
symptom or disease which temporarily resulted from the administration of any research 
procedures associated with the study. The expression “reasonable causal relationship” means 
to convey, in general, that there is evidence or argument to suggest a causal relationship. 
 
6.9 Definition of Serious Adverse Reaction (SAR) 
 

 
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A SAR is an AR that is classed as serious as per the criteria included in section 6.6 of the 
study protocol. 
 
6.10 Definition of Suspected Unexpected Serious Adverse Reaction (SUSAR) 
 
If an SAE is related to the use of a medical device product or taking part in research 
procedures, and is not listed in the study protocol as an expected occurrence, then it is a 
SUSAR.  
 
6.11 Reporting of SUSARs 
 
Research sites will submit SUSARs to the Sponsor. It is the Sponsor’s responsibility to report 
SUSARs to the REC and to disseminate SUSARs to participating sites. Follow-up of patients 
who have experienced a SUSAR should continue until recovery is complete or the condition 
has stabilised.  
 
6.12 Annual reporting 
 
The Annual Progress Report (APR) will be sent by the CI to the Sponsor and REC using the 
NRES template. The APR will be submitted on the anniversary date of the “favourable opinion” 
letter from the REC. A copy of the APR and an associated correspondence with REC will also 
be sent to participating sites. 
 
6.13 Urgent safety measures 
 
The CI may take urgent safety measures to ensure the safety and protection of the clinical trial 
patients from any immediate hazard to their health and safety, in accordance with Regulation 
30. The measures should be taken immediately. In this instance, the approval of the REC prior 
to implementing these safety measures is not required. However, it is the responsibility of the 
CI to inform the Sponsor and the REC (via telephone for discussion with the medical assessor 
at the clinical trials unit) of this event immediately.  
 
The Sponsor has an obligation to inform the REC in writing within 3 days, in the form of a 
substantial amendment. The Sponsor must be sent a copy of the correspondence with regards 
to this matter. 
 
 
7: STATISTICAL CONSIDERATIONS 
 
7.1 Sample size 
 
A maximum of fifty (50) evaluable patients will be recruited to this study. This study is not 
formally powered due to the proof-of-concept nature of the study and broad inclusion criteria. 
However, the number of samples acquired in this study will enable Pear Bio to conduct 
analytical validation on the robustness of its precision medicine platform in brain cancer. 
 
7.2 Statistical analysis 

 
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7.2.1 Primary efficacy analysis 
 
The primary objective of this study does not require patient outcomes. Instead, comparisons 
will be made to quantify the variability of treatment response within and between patient 
tumour samples. As these are purely analytical comparisons, they are tied to laboratory results 
of the Pear Bio test on patient samples. No correlations are made between Pear Bio test 
results and patient outcomes in the primary analysis.  
 
Comparisons of differentiated ex vivo therapeutic response are done on the basis of: 
 
1. Each therapeutic demonstrating its intended mechanism of action based on a before 
and after biomarker measurement of the tumour/blood sample (e.g., IF assay 
measuring the level of target protein for a given targeted drug) 
 
2. Computer vision analysis applied to confocal microscopy images of the tumour cell 
cultures at multiple timepoints, which yields multiple phenotypic metrics of ex vivo 
tumour response, including: 
a. Cell death 
b. Cell migration distance/speed (mean, median, 5% most aggressive cells, etc.) 
c. Immune cell infiltration into the tumour  
 
These analytical measurements are used to conduct the following comparisons: 
 
1. Observing differentiated ex vivo treatment response across the therapies/combos 
tested on each patient’s tumour sample (intra-patient sample comparison) 
a. Ranking of regimen efficacy will be done for each assay metric on a per patient 
basis to determine agreement/disagreement between assay metrics 
b. Calculating the range of response to all tested regimens for each assay metric 
on a per patient basis 
 
2. Observing differentiated ex vivo treatment response levels between the cohort of 
samples collected from patients on a per therapeutic/combo basis (inter-patient 
sample comparison) 
a. Grouping of patient samples based on change in biomarker levels on a per 
regimen basis 
b. Grouping of patient samples based on ex vivo response or resistance at a 
phenotypic level (quantified by computer vision) on a per regimen basis 
c. Comparing the general efficacy of the regimens tested across all patient 
samples using box plots for each phenotypic ex vivo tumour response metric 
of interest 
d. Comparing the general efficacy of the regimens tested across patient samples 
using regimen efficacy ranks on each sample and using Kendall’s W and 
Spearman ranked correlation tests to determine whether some regimens 
consistently outperform others ex vivo (on a given metric of interest) 

 
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e. Comparing the general efficacy of the regimens tested across patient samples 
using Repeated Measures ANOVA for each phenotypic ex vivo tumour 
response metric of interest 
f. Comparing the relative efficacy of any 2 regimens tested across patient 
samples using a paired T-test for each phenotypic ex vivo tumour response 
metric of interest 
 
The primary objective will be met if there are significant variations in intra-patient and inter-
patient sample responses. Due to the heterogeneity of glioma treatment response in the clinic, 
one regimen is not expected to consistently outperform other options across all patient 
samples in the laboratory. That could indicate ex vivo overperformance due to assay 
conditions that require adjustment, either in the biology workflow or response vs resistance 
thresholds, before the assay is used in interventional trials to guide treatment decisions.  
 
7.2.2 Secondary efficacy analysis 
 
Correlating omics biomarkers to ex vivo tumour response 
 
Biomarker expression tied to targeted therapy response (e.g., EGFR to EGFR inhibitors) will 
be correlated to ex vivo tumour response (3D micro-tumour and computer vision platform). 
 
Correlations will be done with various regression models, including: 
 
1. Impact of MGMT methylation on response to treatment 
2. Linear regression between continuous biomarkers and ex vivo tumour response 
metrics (e.g., baseline gene expression vs cell viability after treatment with a targeted 
drug) 
3. (Ordinal) Logistic regression correlating low-high, and potentially low-medium-high,  
biomarker levels against ex vivo treatment sensitivity/resistance based on a 
 
range of image-based metrics, such as 
a. Cell viability 
b. Cell migration speed 
c. Immune cell infiltration into the tumour  
 
This analysis will be used to establish the concordance/discordance between ex vivo 
responses and biomarkers. However, neither the ex vivo responses nor biomarkers will act as 
a ground truth due to the poor correlation between biomarkers and patient response in patients 
with malignant brain tumours.. Discordance (0 < r < 0.7) will be used to identify patient 
populations where Pear Bio may be able to predict response differently from known 
biomarkers. These populations will be considered for future studies to determine whether Pear 
Bio can accurately predict real-world patient response. 
 
7.2.3 Exploratory analyses 
 
Successful tumour cell culture rate from surgical samples 
 
The successful cell culture rate is the percentage of cultures in which ≥70% of viable tumour 
cells plated post-isolation on day 0 are still alive on day 3 in the control hydrogels (cultures 
with no treatment) compared to the number of tumour samples (≥0.4g) arriving 
uncompromised within 24 hours of collection to the Pear Bio laboratory.  
 

 
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Successful immune cell culture rate from blood samples 
 
The successful cell culture rate is the percentage of cultures in which ≥70% of viable immune 
cells plated post-extraction on day 0 are still alive on day 3 in the control well (no treatment) 
compared to the number of blood samples (≥40mL) arriving uncompromised within 24 hours 
of collection to the Pear Bio laboratory.  
 
Correlating ex vivo tumour culture or multi-omic biomarkers to real-world patient outcomes 
 
For any cases where patient outcomes are available or become available prospectively, such 
as recurrence post-surgery, potential predictive or prognostic biomarkers will be identified. As 
this data will be sparse, the analysis will only be used to generate hypotheses for future trials, 
if they present themselves. 
 
Assessing response to experimental therapies 
 
Observing differentiated ex vivo treatment response across the therapies/combos tested on 
each patient’s tumour sample (intra-patient sample comparison) 
a. Ranking of regimen efficacy will be done for each assay metric on a per patient 
basis to determine agreement/disagreement between assay metrics 
b. Calculating the range of response to all tested regimens for each assay metric 
on a per patient basis 
 
 
7.3 Interim analysis and study termination 
 
Interim analysis will be done on laboratory research milestones, such as confirmation of a 
successful hydrogel formulation to culture brain tumour cells after testing the first 5 patient 
samples. This interim analysis will be used to allocate samples for research based on the most 
pressing requirements for ex vivo model validation (e.g., hydrogel formulation, drug dosing, 
etc.).  
 
On recruitment of the first 10 patients, the TMG will meet to assess whether monthly 
recruitment targets are met and to confirm sample quality and successful culture rates upon 
receipt and processing at the Pear Bio lab. The TMG will use the results to determine whether 
to increase accrual up to a maximum of 50 patients. 
 
7.4 End of study definition 
 
The end of the trial is defined as the last patient's last data at a maximum of 12 months post-
surgery (outstanding data sent to Pear Bio or final laboratory readout, whichever happens 
later). In cases of early termination of the trial (e.g., due to slow accrual) or a temporary halt, 
the Sponsor will notify the REC within 15 days of the decision, and a detailed written 
explanation for the termination/halt will be given.  
 
7.5 Handling of missing data 
 
Missing data will be recorded as not available on eCRFs. Missing data points will not be 
imputed in the analysis for that specific endpoint. 

 
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8: DATA HANDLING AND RECORD KEEPING 
 
8.1 Confidentiality 
 
All information generated in the study will be kept strictly confidential. The researchers 
conducting the trial will abide by the Data Protection Act 1998, and the rights the patient has 
under this act. 
 
Parts of the patients’ medical records and the data collected for the trial will be looked at by 
authorised personnel from the Sponsor. It may also be looked at by authorised personnel from 
the patient’s NHS Trust to check that the trial is being carried out correctly. This is clearly 
stated on the consent form. 
 
All the above bodies have a duty of confidentiality to the patient as a research participant and 
nothing that could reveal their identity will be disclosed outside the research site. All data will 
be stored in a locked and dedicated room only accessible by authorised personnel. 
 
8.2 Study documents 
 
All trial related documents should be filed in the Investigator’s Site File (ISF). It should contain 
essential documents as per the contents page provided to the Investigator by the Sponsor. 
The Sponsor will inform the PI and their staff of any updates and forward any relevant 
documentation. It is the participating PI’s responsibility to maintain this file and keep all records 
up to date. 
 
8.3 Data and sample acquisition 
 
 
This trial uses electronic case report forms (eCRFs). Sites will receive training for appropriate 
eCRF completion. The eCRFs will be submitted electronically to the Sponsor and should be 
handled in accordance with the Sponsor’s instructions.  Any data queries arising from initial 
review will be sent to the relevant centre for resolution. 
 
All eCRFs should be completed by designated, trained examining personnel or the study 
coordinator as appropriate. The eCRF should be reviewed and electronically signed and dated 
by the investigator. In addition, at the end of the study, the investigator will receive patient data 
for his or her site that must be kept with the study records.   
 
The Trial Management Group (TMG) reserves the right to amend or add to the eCRFs as 
appropriate. Revised or additional forms should be used by centres in accordance with the 
guidelines provided by the Sponsor. 
 
The PI will be responsible for monitoring the transfer of biological specimens. The Sponsor 
will confirm the receipt of biological specimens. Tracking forms will accompany all sample 
transfers to the Sponsor’s central lab. The clinical site will link with the Sponsor to ensure all 

 
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biological samples are collected and transferred as per the lab manual. All data will be 
handled, computerised and stored in accordance with GDPR.   
 
PPIE work will take place within the host NHS Trust, and any notes will be kept securely by 
the clinical team. Any results will be provided in an anonymised, summarised format only. Staff 
from the Sponsor may be invited to take part in these sessions in order to explain the product 
in detail, but will not be allowed to collect identifiable data on any participants. 
 
8.4 Record retention and archiving 
 
At the end of the trial, all documentation, as defined by GCP, should be stored by each 
individual site’s archiving facility, until notification for destruction from the Sponsor.  The 
location of the archiving facility must be provided to the Sponsor. 
 
The Sponsor will arrange a ‘close out’ visit where all trial documentation will be prepared for 
archiving by that site. Records will be retained at each individual site. All records relating to 
the trial should be stored together, including the ISF. It is the responsibility of the Principal 
Investigator to ensure a full set of records is collated and documented.  
 
In addition, source documentation (medical notes, images, results etc.) should be retained, as 
per Sponsor request, for the duration of the archiving period. 
 
All this information will be stored for a minimum of 25 years. The Sponsor should be contacted 
prior to destruction. 
 
8.5 Compliance 
 
This trial will be conducted in accordance with the principles of Good Clinical Practice (GCP) 
as laid out in the EU directive and The Medicines for Human Use (Clinical Trials) Regulation 
2004, and its amendments.  
 
In addition, Sponsor auditors will be allowed access to eCRFs, source documents, and other 
trial files to evaluate the trial. Audit reports will be kept confidential. 
 
9: STUDY MANAGEMENT 
 
A TMG will be convened and will consist of members of the clinical coordinating centre (CI, 
Trial Coordinator, Clinical Research Fellow) and the Sponsor’s representatives, scientists and 
statistician(s). The role of the TMG will be to monitor all aspects of the conduct and progress 
of the trial, ensure that the protocol is adhered to, and take appropriate action to safeguard 
participants and the quality of the trial itself. The TMG will meet at least twice a year. Final 
decisions about the continuation or termination of the trial are the responsibility of the TMG. 
 
10: CLINICAL GOVERNANCE ISSUES 
 
10.1 Ethical considerations 
 

 
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The trial will be conducted in accordance with ethical principles founded in the Declaration of 
Helsinki. The Research Ethics Committee (REC) will review all appropriate trial documentation 
in order to safeguard the rights, safety and wellbeing of patients. The trial will only be 
conducted at sites where appropriate approval has been obtained. 
 
The Sponsor will inform the REC of any changes to the conduct of the trial and seek approval 
for these changes and any amended patient materials. The Sponsor will maintain an accurate 
and complete record of all written correspondence to and from the REC and will agree to share 
all such documents and reports with the Sponsor. 
 
The informed consent and any other documentation provided to patients will be revised if 
important new information becomes available that is relevant to the subject’s consent. 
Amended documents will be approved by the REC before distribution to patients. 
 
Participation in the PPIE stream will be by invitation, and participants will be provided with 
written information on background, purpose and use of data from the sessions. 
 
10.2 Summary of monitoring plan 
 
Refer to the PEAR-GLIO Monitoring Plan for further details. Monitoring will involve a review of 
the Investigator Site File (ISF), as well as a proportion of Source Data Verification (SDV). This 
will involve direct access by Sponsor representatives (or other parties, see Section 8.1) to 
patient notes at the participating hospital sites, which will include the review of consent forms 
and other relevant investigational reports. Missing data will be sought, unless confirmed as 
not available. During these visits, the site’s activity will be monitored to verify that: 
 
● Source data transcribed onto eCRFs is authentic, accurate, and complete 
● Safety, rights, and well-being of the participants are being protected 
● The study is being conducted in accordance with the currently approved protocol 
● Any other study agreements, GCP, and all applicable NRES requirements are met 
 
10.3 Audit and inspection 
 
This study may be audited by representatives from the Sponsor. The investigator and 
institution will be informed of the audit outcome. Investigators are obliged to cooperate in any 
audit; allowing the auditor direct access to all relevant documents and allocating their time and 
the time of their staff to the auditor to discuss any findings or issues. Audits may occur at any 
time during or after completion of the study. The investigator should notify the Sponsor 
immediately of any other audits/inspections if there are any such plans. 
 
10.4 Reporting of serious breaches in GCP or the trial protocol 
 
All investigators participating in the trial will promptly notify the Sponsor of a serious breach 
(as defined in Regulation 29A of the Medicines for Human Use (Clinical Trials) Regulations 
2004 [Statutory Instrument 2004/1031], as amended by Statutory Instrument 2006/1928) that 
they become aware of. The CI is responsible for notifying the Sponsor within 24 hours of 
becoming aware of a serious breach. 

 
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The Sponsor is responsible, within 7 days of becoming aware of that breach, for notifying the 
REC in writing of any serious breach of: 
 
● The conditions and principles of GCP in connection with the trial; or 
● The protocol relating to that trial, as amended from time to time in accordance with 
regulations 22 to 25. 
 
A “serious breach” is a breach which is likely to affect to a significant degree:  
● The safety or physical or mental integrity of patients in the trial; or  
● The scientific value of the trial. 
 
11: STUDY FINANCES 
 
11.1 Funding sources 
 
This trial is Sponsor-led. Funding is provided by Ourotech Limited (trading as Pear Bio). 
 
11.2 Patient expenses/payments 
 
The Sponsor will compensate study participants for any additional visits related to participation 
in this trial (i.e., visits outside standard care). This will only cover study participants in the UK, 
and only for UK domestic travel. 
 
12: SPONSORSHIP AND INDEMNITY 
 
Dr. Matthew Williams is the Chief Investigator. Ourotech Limited (trading as Pear Bio) is 
sponsoring the study. Indemnity for participating sites is provided by the Sponsor. 
 
13: PUBLICATION POLICY 
 
This study is sponsored by Ourotech Limited (trading as Pear Bio). The data collected in this 
study will not be used to licence/register any pharmaceuticals. Authorship of the final 
manuscript(s), interim publications, or abstracts will be decided according to active 
participation in statistical design, TMG, accrual of eligible patients and statistical analysis.  
 
Contributing centres (and participating investigators) will be acknowledged in the final 
manuscript.  Representatives of the Sponsor will be added, as appropriate, as co-authors. No 
participant may present data from their centre separately from the rest of the study results, 
unless they receive written approval from the TMG and the Sponsor. The publication policy 
will adhere to the contractual agreement between the Sponsor and its collaborators. 
 
14: REFERENCES 
 
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https://www.cancerresearchuk.org/health-professional/cancer-statistics/statistics-by-cancer-
type/brain-other-cns-and-intracranial-tumours 

 
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2. Siegel RL, Miller KD, Fuchs HE, Jemal A. Cancer statistics, 2022. CA Cancer J Clin. 2022 
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3. Tan AC, Ashley DM, López GY, Malinzak M, Friedman HS, Khasraw M. Management of 
glioblastoma: State of the art and future directions. CA Cancer J Clin. 2020 Jul;70(4):299–312.  
4. van den Bent MJ, Tesileanu CMS, Wick W, Sanson M, Brandes AA, Clement PM, et al. Adjuvant 
and concurrent temozolomide for 1p/19q non-co-deleted anaplastic glioma (CATNON; EORTC 
study 26053-22054): second interim analysis of a randomised, open-label, phase 3 study. Lancet 
Oncol. 2021 Jun;22(6):813–23.  
5. Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, et al. Effects of 
radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in 
glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet 
Oncol. 2009 May;10(5):459–66.