Arrowhead Pharmaceuticals : EASL 2023 JNJ-3989 REEF-2

Association of Post-treatment Virologic Relapse and Biochemical Flares With HBV Serum Biomarkers in Long-term Virologically Suppressed HBeAg negative Patients Stopping NA Treatment: Exploratory Analyses From the Control Arm of the REEF-2 Study

Florian van Bömmel,1,* Thierry Verbinnen,2 Kosh Agarwal,3 Thomas Vanwolleghem,4 Pietro Lampertico,5,6 Maria Buti,7 Ewa Janczewska,8 Marc Bourliere,9 John Jezorwski,10 Kathleen Donohue,11 Gloria Kim,12 Thomas N. Kakuda,13 Sandra De Meyer,2 Adam Bakala,2 Oliver Lenz,2 Michael Biermer2

1Division of Hepatology, Department of Medicine II, Leipzig University Medical Center, Leipzig, Germany; 2Janssen Pharmaceutica NV, Beerse, Belgium; 3Institute of Liver Studies, King's College Hospital, Denmark Hill, London, UK; 4University of Antwerp, Faculty of Medicine & Health Sciences, Laboratory of Experimental Medicine & Pediatrics, Viral Hepatitis Research Group, Antwerp, Belgium;
5Foundation IRCCS Ca' Granda Ospedale Maggiore Policlinico, Division of Gastroenterology and Hepatology, Milan, Italy; 6CRC "A. M. and A. Migliavacca" Center for Liver Disease, Department of Pathophysiology and Transplantation, University of Milan, Milan, Italy; 7Hospital General Universitari Vall Hebron and CIBER EHD del Instituto Carlos III, Barcelona, Spain; 8Medical University of Silesia in Katowice,	*Presenting author.
Faculty of Public Health in Bytom, Bytom, Poland; 9Hôpital Saint-Joseph, Department of Hepatology and Gastroenterology, Marseille, France; 10Janssen Research & Development, LLC, Titusville, NJ, USA; 11Cytel Inc., Waltham, MA, USA; 12IQVIA, Durham, NC, USA; 13Janssen Research & Development, LLC, Brisbane, CA, USA.

SAT-154

Key Findings

In the NA only arm,

off-treatment virologic

> relapses and ALT flares were

experienced by 27 (66%)

Introduction

HBcrAg

- 7 (50%) had detectable off-treatment HBV DNA levels varying between >200 and ≤2,000 IU/mL. 5 (36%) additional

patients had detectable off-treatment HBV DNA >200 IU/mL with single peak values >2,000 IU/mL (n = 4) and

Figure 4. (A) Peak ALT and (B) peak HBV DNA during follow-up versus EOT anti-HBc IgG levels.

• • The phase 2b REEF-2 study (ClinicalTrials.gov Identifier: NCT04129554) assessed the efficacy and safety of the combination of short-interfering RNA JNJ-73763989(JNJ-3989), capsid assembly modulator JNJ-56136379(JNJ-6379; bersacapavir), and nucleos(t)ide analogue (NA) compared to NA only in long-term, virologically suppressed hepatitis B e antigen (HBeAg) negative patients with chronic hepatitis B (CHB)1
  • • All treatments were discontinued in both arms at Week 48, followed by 48 weeks of follow-up1
    • In the NA arm, a higher rate of virologic relapse, post-treatment alanine aminotransferase (ALT) flares, and NA retreatment with a lower rate of sustained off-treatment hepatitis B surface antigen (HBsAg) and hepatitis B virus (HBV) DNA suppression were observed compared to the JNJ-3989 arm1
• While several recent studies have demonstrated that stopping NA treatment can result in HBsAg seroclearance in some patients with CHB,2,3 almost all patients who stop NA treatment experience an off-treatment transient virologic relapse, which can be associated with severe ALT increases in some patients
• Varying levels of end of treatment (EOT) viral parameters, such as HBsAg, hepatitis B core-related antigen (HBcrAg), HBV RNA, and hepatitis B core antibody (anti-HBc), have been demonstrated to be associated with the risk of virologic relapse and biochemical (ALT) flares after stopping NA treatment4-8

Objective

• To characterize post-treatment virologic relapses and biochemical flares in the NA arm of the REEF-2 study and assess their association with EOT HBV serum markers

Methods

•	"Standard" testing at the central laboratory showed that 24/41 (58.5%) patients had HBcrAg 10 U/mL)
	at EOT (Table 1)
•	For exploratory retesting, all EOT samples with a result of HBcrAg <3.0 log10 U/mL were considered:
	- All 24 reanalyzed samples were confirmed to have HBcrAg <3.0 log10 U/mL in the reanalysis, and 20 samples were
	determined to have HBcrAg <2.6 log10 U/mL (ie, Table 1)

Anti-HBc IgG

• The mean (range) anti-HBc IgG level at EOT was 1,366 (7-11,000) IU/mL, and 23/41 (56%) patients had EOT anti-HBc IgG levels <300 IU/mL (Table 1)

Table 1. REEF-2 NA Control Arm Patient Disease Characteristics at EOT

			"Standard" assay*,†						Exploratory analysis/assay*,†
HBeAg negative		41 (100)						-
Mean (range) HBsAg, log10 IU/mL		3.42 (1.8-4.7)						-
HBsAg <1,000 IU/mL		13 (31.7)‡						-
HBV DNA <>§		41 (100)						-
									HBcrAg <2.6 log10 U/mL (ie,	20 (48.8)
HBcrAg <>�		24 (58.5)	HBcrAg ≥2.6 to <3.0 log10	U/mL	4 (9.8)
									(
													≥LLOQ	11 (26.8)
									HBV RNA assay,**
											2 (4.9)
									0.2 mL
HBV RNA <>	¶				39 (97.5)#						5 (12.2)
(n = 40)									≥LLOQ	22 (53.6)
									HBV RNA assay,**
											0
									0.6 mL

	>20,000 IU/mL (n = 1)
- Few had detectable HBV RNA and HBcrAg levels, mostly at single time points
-	1 (7%) had transiently detectable HBeAg, and none had increases in HBsAg
- None were retreated with NA, and 12 (86%) had HBV DNA >LLOQ and <2,000 IU/mL, and 2 had HBV DNA
	Follow-up Week 48

Association of Post-treatment Virologic Relapse and Biochemical Flares With EOT HBV Serum Biomarkers

HBV RNA, HBcrAg, HBsAg, and HBV DNA

• A similar proportion of patients with HBV RNA detectable and TND at EOT had virologic relapse or biochemical flares (Table 2)
• A higher frequency of virologic relapse with peak HBV DNA >100,000 IU/mL and biochemical flares with peak
  ALT ≥10 × ULN was observed in patients with detectable HBcrAg at EOT compared to those with HBcrAg TND (Table 2)
• A higher frequency of virologic relapse with peak HBV DNA >100,000 IU/mL and biochemical flares was observed in patients with HBsAg <1,000 IU/mL at EOT compared to those with HBsAg ≥1,000 IU/mL (Table 2)
• A similar proportion of patients with HBV DNA

Anti-HBc IgG

•

Patients with anti-HBc IgG titers <300 IU/mL at EOT had a significantly higher frequency of virologic relapse with peak

HBV DNA >100,000 IU/mL or biochemical flares than patients with EOT anti-HBc IgG titers ≥300 IU/mL (Table 2; Figure 4).

Notably, none of the 11 patients with anti-HBc IgG titers ≥300 IU/mL at EOT had peak HBV DNA >100,000 IU/mL or

peak ALT ≥10 × ULN (Table 2; Figure 4)

•

For prediction of (a) the absence of severe biochemical flare (ie, peak ALT level ≥10 × ULN) and (b) the absence of

virologic relapse with peak HBV DNA >100,000 IU/mL, patients with EOT anti-HBc IgG ≥300 IU/mL had:

- (a) 58% sensitivity, 100% specificity; positive predictive value (PPV) of 100%, negative predictive value (NPV) of 43%

- (b) 60% sensitivity, 100% specificity; PPV of 100%, NPV of 48% (Figure 5)

A.

Peak ALT (U/L) during follow-up

4,000

1,000

400

100

40

10

							B.
				IU/mL
				300				1 × 109
								1 × 108
							-up
							follow	1 × 107
							during	1 × 106
							(IU/mL)
							100,000
							DNAHBV
							10,000
							Peak	1,000
								100
								10
10	100	1,000	10,000

Anti-HBc IgG result (IU/mL)

300 IU/mL

10

100

1,000

10,000

Anti-HBc IgG result (IU/mL)

Study Design

• The current analysis assessed the post-treatment responses of patients in the NA (control) arm of the REEF-2 study (Figure 1)

Figure 1. Study design.

Randomized, double-blind, multicenter,				Study intervention	Primary		End
placebo-controlled study				stopped (including NA)	endpoint‡		of study
NA suppressed/HBeAg negative
CHB with NA treatment 2 years
ALT <2.0 × ULN		JNJ-3989* + JNJ-6379† + NA (n = 85)
HBV DNA <60 IU/mL				Follow-up
HBsAg >100 IU/mL at screening			(active arm)
Non-cirrhotic (fibrosis stage F0-F2)	JNJ-3989 placebo + JNJ-6379 placebo + NA
NA = ETV/TAF/TDF according to label		Follow-up
	(n = 45) (control arm)
7 countries in Europe
Week	0	12	24	36	48	F12	F24	F36	F48
No. of patients per analysis visit	130	119	118	117	117	120	119	120	121

ETV, entecavir; F, follow-up; LLOQ, lower limit of quantification; PO, oral; SC, subcutaneous; TAF, tenofovir alafenamide; TDF, tenofovir disoproxil; ULN, upper limit of normal. *200 mg SC every 4 weeks. †250 mg PO daily. ‡HBsAg seroclearance (HBsAg

NA Retreatment Criteria

• Patients needed to restart NA treatment immediately in the case of signs of decreasing liver function or (after protocol amendment) HBV DNA >100,000 IU/mL (irrespective of confirmation and/or ALT increase)
• Restart of NA was to be considered in the case of post-treatment HBeAg seroreversion, confirmed (≥4 weeks apart) post-treatment increases in HBV DNA >20,000 IU/mL, or HBV DNA >2,000 IU/mL in combination with ALT >5 × ULN

Definitions

• Virologic relapse was defined as confirmed (ie, 2 consecutive visits) HBV DNA >2,000 IU/mL off treatment
• Biochemical flare was defined as confirmed ALT and/or aspartate transaminase (AST) ≥3 × ULN and ≥3 × nadir off treatment

Assays

•	Baseline and post-baseline ALT, HBV DNA, HBeAg, and HBsAg levels were assessed at a central laboratory (Labcorp)
	during the conduct of this study using "standard" assays. "Standard" HBV RNA levels were assessed at DDL Diagnostic
	Laboratory (The Netherlands) using a validated quantitative reverse transcriptase-polymerase chain reaction with
	a limit of detection (LOD) of 310 copies/mL.9 "Standard" HBcrAg levels were determined at Labcorp using the
•	Lumipulse® platform (Fujirebio) with an LLOQ of 3.0 log10 U/mL
Exploratory analyses of the selected set of samples were performed post hoc and included reanalysis of serum
	HBcrAg and HBV RNA levels and analysis of anti-HBc immunoglobulin G (IgG)
	- HBcrAg was reanalyzed at DDL Diagnostic Laboratory using the same "standard" assay (Fujirebio). In contrast to
	the reporting by the central laboratory, HBcrAg values 10 U/mL were considered. Samples with

			1 (2.4)
ALT <1 × ULN	38 (92.7)	-
	-	Mean (range) anti-HBc IgG, IU/mL	1,367 (7-11,000)

"Standard" assays were run at central laboratories during the study; exploratory assays and analyses were performed post hoc on the selected set of samples. *Values are n (%) unless otherwise noted. †The denominator is the overall population (N = 41). ‡1 patient had HBsAg <100 IU/mL at EOT. §HBV DNA LLOQ = 15 U/mL. �HBcrAg LLOQ = 3.0 log10 U/mL. ¶HBV RNA LOD = 310 copies/mL. #The 1 patient with HBV RNA >LOD in the "standard" assay also had detectable and quantifiable HBV RNA in the exploratory assay. **HBV RNA 0.2 mL (n = 18) and 0.6 mL (n = 23) sample input assay LLOQ = 22 and 11 copies/mL, respectively.

Relationship Between Peak HBV DNA Levels During Virologic Relapse and Magnitude of Off-treatment Biochemical Flares

• Peak HBV DNA levels during virologic relapse were associated with the magnitude of off-treatment ALT flares (R = 0.83; P<0.01; Figure 2)
• 10/11 (91%) patients with virologic relapse and confirmed peak HBV DNA >100,000 IU/mL had peak ALT levels ≥10 × ULN
• • The 1 patient who did not have ALT ≥10 × ULN had a peak ALT level ~9 × ULN
  • 1 other patient had virologic relapse but with an unconfirmed (ie, at a single time point) peak HBV DNA value >100,000 IU/mL. This (male) patient had only limited elevation of ALT levels (peak ALT of 70 U/L)
• None of the 16 patients with virologic relapse and confirmed peak HBV DNA levels >2,000 to ≤100,000 IU/mL had peak ALT levels ≥10 × ULN
• 3 patients had off-treatment ALT flares (2 males with peak ALT ~7 × ULN and 1 female with peak ALT ~4 × ULN) without virologic relapse (peak HBV DNA levels of 54, 532, and 1,620 IU/mL, respectively)

Figure 2. Relationship between peak HBV DNA levels and peak ALT levels during follow-up.

follow-up	4,000			2,000IU/mL	100,000IU/mL
1,000
during	400							10 ULN (M)
						10 ULN (F)
(U/L)ALT
100							3 ULN (M)
							3 ULN (F)
Peak	40
	10							R = 0.83; P<0.01
	10	100	1,000	10,000	100,000 1 × 106	1 × 107	1 × 108	1 × 109

Peak HBV DNA (IU/mL) during follow-up

Diamonds and circles represent data for female and male patients, respectively. Data in yellow indicate patients who restarted NA treatment. Solid and dashed

• With the addition of HBcrAg, the accuracy measurements improved compared to the model with anti-HBc IgG alone.

Using an example for the Firth's logistic regression, a patient with anti-HBc IgG ≥300 IU/mL and HBcrAg TND at EOT

had a predicted probability of 0.009 to have a peak ALT level ≥10 × ULN and 0.01497 to have peak HBV DNA >100,000 IU/mL

(vs 0.026 with anti-HBc IgG alone; Figure 5)

Univariate and Multivariate Logistic Regression of EOT Virologic Parameters

• EOT HBsAg, HBcrAg, and anti-HBc IgG each showed univariate association with the outcome variable absence of peak ALT ≥10 × ULN during follow-up (Table 3)
• HBsAg was no longer significant when adjusting for HBcrAg and anti-HBc IgG in a multivariate model. When adjusting for anti-HBc IgG, HBcrAg was marginally significant (Table 3). The odds ratio (OR) for not having a peak ALT level ≥10 × ULN for those with EOT anti-HBc IgG ≥300 IU/mL was 21.402 (95% confidence interval [CI], 2.187-2,902.034;P = 0.0457). For those with EOT HBcrAg 1.161-80.166;P = 0.0564)

Table 2. Proportion of REEF-2NA Control Arm Patients With Post-treatmentVirologic Relapse and Biochemical Flare by EOT Virologic Parameters

		Virologic relapse	Biochemical flare
		(confirmed HBV DNA >2,000 IU/mL	(ALT ≥3 × ULN)
		Any virologic	Peak HBV DNA		Peak ALT
EOT variables/type of NA	N	flare	>100,000 IU/mL	Any ALT flare	≥10 × ULN
Patients with EOT data who	41	27 (65.9)	11 (26.8)	16 (39.0)	10 (24.4)
entered follow-up, n (%)
HBV RNA
Detectable*	35	23 (65.7)	9 (25.7)	13 (37.1)	9 (25.7)
TND†	6	4 (66.7)	2 (33.3)	3 (50.0)	1 (16.7)
P value‡		1.0000	0.6514	0.6624	1.0000
HBcrAg	21	17 (81.0)	9 (42.9)	10 (47.6)	9 (42.9)
Detectable*
TND†	20	10 (50.0)	2 (10.0)	6 (30.0)	1 (5.0)
P value‡		0.0516	0.0325	0.3408	0.0089
Anti-HBc IgG
<300 IU/mL	23	16 (69.6)	11 (47.8)	12 (52.2)	10 (43.5)
≥300 IU/mL	18	11 (61.1)	0	4 (22.2)	0
P value‡		0.7417	0.0008	0.0626	0.0021
HBsAg
<1,000 IU/mL§	13	9 (69.2)	6 (46.2)	8 (61.5)	6 (46.2)
≥1,000 IU/mL	28	18 (64.3)	5 (17.9)	8 (28.6)	4 (14.3)
P value‡		1.0000	0.0727	0.0835	0.0485

Bold values are significant. *‡2-sidedP value from Fisher's exact test. No adjustments for multiplicity were performed. Statistical testing was performed to compare the exploratory biomarker classifications among the patients who had any virologic flare (yes vs no), off-treatment virologic flare of >100,000 IU/mL (yes vs no), any ALT flare (yes vs no), and peak ALT ≥10 × ULN in the follow-up phase (yes vs no). §Includes 1 patient with

EOT HBsAg <100 IU/mL.

Circles and triangles represent data for male and female patients, respectively. Data in yellow indicate patients who restarted NA treatment. Solid and dashed horizontal red lines in panel A represent ALT values of 10 × ULN for females (340 U/L) and males (430 U/L), respectively. The solid black lines are linear regression trend lines.

Table 3. Logistic Regression Model for Not Having a Biochemical Flare With Peak ALT ≥10 × ULN During Follow-upby EOT Virologic Parameters

		Univariate model		Multivariate model
EOT variables	OR (95% CI)*	P value	OR (95% CI)*	P value
HBV DNA	1.381	(0.043-97.797)	0.8640	-	-
HBV RNA	1.731	(0.235-35.495)	0.6368	-	-
HBcrAg	14.248 (2.261-280.182)	0.0174	7.150 (1.161-80.166)	0.0564
Anti-HBc IgG ≥300 IU/mL†	28.775 (3.204-3,820.88‡)	0.0282	21.402 (2.187-2,902.034‡)	0.0457
HBsAg <1,000 IU/mL	0.194 (0.039-0.862)	0.0347	-	-

Bold values are significant. *Profile likelihood method. †Logistic regression using Firth's bias correction. ‡The R software package calculated the upper CIs to be 3,820.88 and 2,902.034.

Figure 5. Sensitivity and specificity of anti-HBcIgG and HBcrAg for predicting (A) the absence of biochemical flare with peak ALT ≥10 × ULN and (B) virologic relapse with peak HBV DNA >100,000 IU/mL.

	Anti-HBc IgG	Anti-HBc IgG and HBcrAg
A.	ROC for peak ALT ≥10 × ULN	B.	ROC for peak HBV DNA >100,000 IU/mL

	1.0							1.0
Sensitivity	0.8						Sensitivity	0.8
0.6						0.6
	0.4							0.4
	0.2							0.2
	0							0
	1.0	0.8	0.6	0.4	0.2	0		1.0	0.8	0.6	0.4	0.2	0
			Specificity						Specificity
	Anti-HBc IgG + HBcrAg vs anti-HBc IgG alone:			Anti-HBc IgG + HBcrAg vs anti-HBc IgG alone:
	sensitivity = 0.81 vs 0.58; specificity = 0.90 vs 1.00;		sensitivity = 0.80 vs 0.60; specificity = 0.82 vs 1.00;
	PPV = 0.96 vs 1.00; NPV = 0.60 vs 0.43; AUC = 0.88 vs 0.79		PPV = 0.92 vs 1.00; NPV = 0.60 vs 0.48; AUC = 0.86 vs 0.80

AUC, area under the curve; ROC, receiver operating characteristic.

an HBcrAg result <2.6 log10 U/mL were categorized as

result ≥2.6 to <3.0 log U/mL were categorized as

horizontal lines represent ALT values of 3 × ULN (green) and 10 × ULN (red) for females (102 and 340 U/mL) and males (129 and 430 U/L), respectively. The solid black line is a linear regression trend line.

Figure 3. Individual HBV DNA, ALT, HBsAg, HBV RNA, HBeAg, HBcrAg, and anti-HBc profiles by peak HBV DNA value during follow-up.

-	10
Exploratory HBV RNA analysis was performed at bioMONTR® (United States) using the Abbott HBV RNA assay.10
	This assay is available in 2 versions according to the sample input volume being either 0.2 mL (LLOQ of
	22 copies/mL) or 0.6 mL (LLOQ of 11 copies/mL)
- Anti-HBc IgG levels were analyzed at DDL Diagnostic Laboratory using the Lumipulse® (Fujirebio) anti-HBc assay11

Results

Patients

• 41/45 patients enrolled in the REEF-2 NA control arm were included in this analysis (2 patients discontinued study treatment at Week 2 and did not enter follow-up, 1 patient discontinued study treatment at Week 8 and entered follow-up but continued NA treatment, and 1 patient completed study treatment but did not enter follow-up)
• All patients were long-term virologically suppressed (>2 years) and HBeAg negative at screening. The majority were male (64%); the mean age was 47.4 years; and, while the study was based in Europe, ~18% of patients were Asian. The mean duration of NA treatment at study entry was 8.1 (range: 2.2-17.4) years

Exploratory Assay Analyses

HBV RNA

•	EOT "standard" assay HBV RNA data were available for 40/41 patients, and 39/40 (97.5%) had HBV RNA
•	"standard" assay (LOD = 310 copies/mL9) at EOT (Table 1)
All EOT samples (N = 41) were reanalyzed with the Abbott HBV RNA assay:
	- 23 samples were reanalyzed using a sample input volume of 0.6 mL. The other 18 samples were reanalyzed using a
	sample input volume of 0.2 mL due to limited available sample volume

Kinetics of Other Viral Markers During Virologic Relapse

•	Virologic relapse was generally associated with parallel increases in other viral markers, especially among patients with
	high peak HBV DNA levels during follow-up (Figure 3)
•	Among patients with virologic relapse and confirmed peak HBV DNA levels >100,000 IU/mL (n = 11), during
	virologic relapse:
	- 11 (100%) had confirmed HBV RNA increases ≥2 log10 cp/mL from EOT levels (all had HBV RNA
	assay at EOT)
	- 11 (100%) had >1.0 log10 IU/mL increase in anti-HBc IgG levels from EOT
	- 8 (73%) had confirmed HBcrAg increase ≥1.0 log10 U/mL from EOT levels (including 4 patients who had EOT
	HBcrAg levels <>
	- 8 (73%) had transient HBsAg increase ≥0.5 log10 IU/mL from EOT levels (5 had ≥1 log10 IU/mL increase)
	- 4 (36%) had transient detectable HBeAg levels during follow-up (maximum increases in HBeAg levels ranged
	from 0.5 to 3.9 log10 IU/mL)
	- 10 (91%) restarted NA treatment, which resulted in declines in virologic markers in all patients
•	ƒ 7 and 8 had HBV DNA <_uln29_ at="">Follow-up Week 48, respectively
Among patients with virologic relapse and confirmed peak HBV DNA levels >2,000 to ≤100,000 IU/mL (n = 16),
	during off-treatmentfollow-up:
	- Increases in HBV RNA and HBcrAg levels were more modest, less frequent, and, in the majority of patients,
	at single time points (ie, blips)
	- No associated increases in HBsAg levels and no transient detectable HBeAg levels were observed
	- 2 (13%) restarted NA treatment and had HBV DNA Follow-up Week 48
	- All 14 others who were not retreated with NA still had detectable HBV DNA levels at Follow-up Week 48

Virologic relapse	and peak HBV	DNA >100,000 IU/mL	(n = 11)
Virologic relapse and	peak HBV DNA >2,000	to ≤100,000 IU/mL	(n = 16)
No virologic	relapse	(n = 14)

10

9

8

7

6

5

4

3

2

1

10

9

8

7

6

5

4

3

2

1

10

9

8

7

6

5

4

3

2

1

HBV DNA

(log10 IU/mL)

2,000

1,800

1,600

1,400

1,200

1,000

800

600

400

200

0

0 100 200 300 400 500 600 700

350

300

250

200

150

100

50

0

0 100 200 300 400 500 600 700

350

300

250

200

150

100

50

0

0 100 200 300 400 500 600 700

ALT

(U/L)

5

4

3

2

1

0

-1

0 100 200 300 400 500 600 700

5

4

3

2

1

0

-1

0 100 200 300 400 500 600 700

5

4

3

2

1

0

-1

0 100 200 300 400 500 600 700

HBsAg

(log10 IU/mL)

9

8

7

6

5

4

3

*2

0 100 200 300 400 500 600 700

9

8

7

6

5

4

3

2

0 100 200 300 400 500 600 700

9

8

7

6

5

4

3

2

0 100 200 300 400 500 600 700

HBV RNA

(log10 copies/mL)

4

3

2

1

0

-1

0 100 200 300 400 500 600 700

4

3

2

1

0

-1

0 100 200 300 400 500 600 700

4

3

2

1

0

-1

0 100 200 300 400 500 600 700

HBeAg

(log10 IU/mL)

10

9

8

7

6

5

4

3

0 100 200 300 400 500 600 700

10

9

8

7

6

5

4

3

0 100 200 300 400 500 600 700

10

9

8

7

6

5

4

3

0 100 200 300 400 500 600 700

HBcrAg	Anti-HBc IgG
(log10 U/mL)	(IU/mL)

4

3

2

1

0

100

200

300

400

500

600

700

0

100

200

300

400

500

600

700

10,000

1,000

								100
								10
								1
0	100	200	300	400	500	600	700	0	100	200	300	400	500	600	700

10,000

1,000

100

								10
								1
0	100	200	300	400	500	600	700	0	100	200	300	400	500	600	700

- After reanalysis with the Abbott HBV RNA assay, 6/41 (15%) patients had HBV RNA

and 1/23 (4%) patients were analyzed using sample input volumes of 0.2 and 0.6 mL, respectively (Table 1)

(8 with HBV DNA levels ≥2,000 IU/mL)

• Among patients with no virologic relapse (n = 14), during off-treatmentfollow-up:

The color of the lines indicates the level of virologic relapse for individual patients. Red: HBV DNA >100,000 IU/mL; blue: >2,000 to ≤100,000 IU/mL; gray: no virologic relapse. Time points shown in yellow indicate time points after the start of NA retreatment. Data shown are from "standard" assays. X-axis values represent days after the start of study treatment. Vertical lines indicate EOT (Week 48). Virologic relapse is defined as confirmed off-treatment HBV DNA levels >2,000 IU/mL. *1 patient experienced severe HBV reactivation with subacute liver failure that led to transplantation at Follow-up Week 13.

and 16 (39%) patients,

respectively, after NA

treatment discontinuation

Peak HBV DNA levels

during virologic relapse were

> associated with the magnitude

of ALT flares

Assessment of EOT viral

markers and off-treatment

> response showed:

- Virologic relapse and

ALT flares were observed

at a similar rate for

patients regardless

of the detectability

of HBV RNA

- Virologic relapse and

biochemical flares with

high peak HBV DNA or ALT

values were more frequent

in patients with detectable

HBcrAg and HBsAg

<1,000 IU/mL at EOT

- None of the 18 patients with

anti-HBc IgG ≥300 IU/mL

at EOT had peak HBV DNA

>100,000 IU/mL or peak

ALT ≥10 × ULN

Conclusions

>	In patients discontinuing NA
treatment, the increases in
HBV DNA during virologic
	relapse correlated with peak
	levels of ALT
	EOT anti-HBc IgG levels
	≥300 IU/mL were strongly
> associated with the absence of
	high peak HBV DNA virologic
	relapse (>100,000 IU/mL) and

References			Acknowledgments	Disclosures
1.	Agarwal K, et al. Presented at: AASLD The Liver Meeting;	6.	Chi H, et al. Clin Gastroenterol Hepatol. 2019;17(1):182-191.e1.	This study was supported by Janssen Research & Development, LLC. Editorial support was provided by Kim Caldwell, PhD, of Lumanity Communications Inc., and was funded by Janssen Global Services, LLC.	FvB consults for, is on the speakers bureau for, and received grants from Gilead, Bristol Myers Squibb, and Roche; and has scientific collaboration with Roche and Janssen. T Verbinnen, JJ, TNK, SDM, AB, OL, and M Biermer are employees of Janssen and may hold stock in Johnson & Johnson. KA served on the advisory board/speakers
	November 4-8, 2022; Washington, DC, USA.	7.	Adraneda C, et al. J Hepatol. 2023;78(4):731-741.		bureau for Arbutus, Assembly, Aligos, Biotest, Bluejay, Drug Farm, GSK, Gilead, Janssen, Intercept, Merck, Roche, Sagimet, Sobi, and Vaccitech; and received research support from Gilead and Abbott. T Vanwolleghem receives grants from Gilead and Bristol Myers Squibb; and serves as a consultant for Janssen, Gilead, and AbbVie.
2.	Berg T, et al. J Hepatol. 2017;67(5):918-924.	8.	Liu J, et al. Hepatology. 2019;70(3):1045-1055.		PL serves on the advisory board/speakers bureau for Bristol Myers Squibb, Roche, Gilead, GSK, AbbVie, MSD, Arrowhead, Alnylam, Janssen, Spring Bank, MYR, Eiger BioPharmaceuticals, Antios Therapeutics, Aligos, and Vir Biotechnology. M Buti received grants from Gilead; and gave sponsored lectures for Gilead and AbbVie.
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high peak ALT flares (≥10 × ULN);

this association was weaker for

EOT HBcrAg and HBsAg levels

and absent for HBV RNA

Presented at the European Association for the Study of the Liver (EASL) International Liver Congress™; June 21-24, 2023; Vienna, Austria.