Genprex : AACR 2026 Poster - Quaratusugene Ozeplasmid Mediated TUSC2 Upregulation In EML4-ALK Bearing Non-small Cell Lung Carcinoma Induces Apoptosis And Is Highly Effective In Preclinical Studies

Published: 2026-04-19 18:20:05 ET GNPX

Published on 04/19/2026 at 06:20 pm EDT

‌Quaratusugene Ozeplasmid Mediated TUSC2 Upregulation In EML4-ALK Bearing Non-small Cell Lung Carcinoma Induces Apoptosis And Is Highly Effective In Preclinical Studies

Ananya Banerjee1, Neeke Busette1, Xu Cheng1, Kerslee Kohagen1, Liwei Bao1, Lluis Lopez-Barcons1, Rachel Sexton 2, Ross Camidge3, Sharon Pine3, Mark S. Berger4, Matthew B. Soellner2, Angel Qin1, Sofia D. Merajver1*, Nathan M. Merrill1*

1 Department of Internal Medicine, University of Michigan, Ann Arbor, MI, 2 Departments of Medicinal Chemistry and Chemistry, University of Michigan, Ann Arbor, MI,

3 Department of Medicine - Medical Oncology, University of Colorado Anschutz, Aurora, CO, 4 Genprex, Inc. Austin, TX, * Equal contribution

CUTO 8

CUTO 9

CUTO 29.1

✱✱

400

Cleaved

300

150

✱✱✱✱

TUSC2 - - + - - + - - +

✱✱✱✱

200

CUTO 8

CUTO 9

CUTO 29.1

400

✱✱✱✱

✱

✱✱

24 48 72 96 120 144

hours

✱

✱✱✱

BNP + - - + - - + - -

✱✱✱✱

200

- + - - + - - + -

✱✱✱✱

100

200

CUTO 8

CUTO 29.1

✱✱✱✱

Cmax Start low

Start IC50

NCI-H2228

✱✱✱✱

BNP+DMSO QO+DMSO

BNP+Alectinib QO+Alectinib

BNP QO

TUSC2

✱✱✱

Fig 4: QO in combination with Alectinib A,C, can reduce cell viability, B,D,

increase caspase 3/7 activity E,F, inhibit cell proliferation in ALK+ cells that are resistant to Alectinib. QO with Alectinib can also further increase G-J, the expression of TUSC2 and cleaved caspase 3 indicating increased apoptosis and the expression of ʏH2AX, indicating increased DNA damage. Cells were transfected with QO followed by treatment with Alectinib at 10M for 24 hours. Data analyzed using t-test. ****:p<0.0001, ***:0.0001≤p≤0.001, **:0.001≤p≤0.01.

✱✱✱

600

✱✱✱✱

600

✱✱✱✱

NCI-H2228

NCI-H2228 Cmax

400 ✱✱✱✱

✱✱✱✱

300

✱✱✱✱

400

✱✱✱✱

✱✱✱

✱✱

300

200

✱✱✱

100

- - +

- + -

+ - -

TUSC2

400

ALK167-T-01

3000

2000

1000

ALK192-P-01

300 ✱✱✱✱

ALK167-T-01

✱

✱✱

150

Cl BNP

200

100

eaved

PARP

100

Tubulin

TUSC2

Tubulin yH2AX

Tubulin

cleaved caspase 3

Tubulin

BNP QO TUSC2

caspase 3/7 activity

BNP QO TUSC2

caspase 3/7 activity

Fig 2: QO mediated overexpression of TUSC2 A,G, increases the caspase 3/7

activity in ALK+ NSCLC cell lines J,L-M, and in PDOs. B,H,K, increases the expression of cleaved caspase 3 and cleaved PARP. C,I, reduces colony formation ability and D-F, increases DNA fragmentation. Cells were transfected with QO for 48 hrs. Analyzed using t-test. ****:p<0.0001,***:0.0001≤p≤0.001,

**:0.001≤p≤0.01,*:0.01≤p≤ 0.05, ns: p ≥ 0.05.

Caspase 3/7 activity

NCI-H2228

Cmax Start low

TUSC2

Tubulin

100

Caspase 3/7 activity

QO+DMSO

QO+Alectinib

100

caspase 3

Tubulin

200

BNP+DMSO

BNP+Alectinib

✱✱✱✱

NCI-H2228 Cmax

✱✱✱✱

NCI-H2228 Start low

✱✱✱✱

NCI-H2228 Start IC50

✱✱✱✱

Caspase 3/7 activity

200000

150000

100000

50000

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2000

✱✱✱✱

ALK192-P-03

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BNP QO TUSC2

ALK167-T-01

✱✱

CUTO 8

CUTO 9

CUTO 29.1

✱✱✱✱

80000

60000

40000

20000

BNP QO TUSC2 BNP QO TUSC2

Fig 1: A-C, TUSC2 is significantly overexpressed in EML4-ALK+ NSCLC cell lines and D-F, organoids derived from patient tissue (T) and pleural effusion

(P) on transfection with QO for 48 hours. Data analyzed using t-test.

**:0.001≤p≤0.01, ****:p < 0.0001.

QO mediated TUSC2 overexpression can induce apoptosis in EML4-ALK+ NSCLC cell lines

Fig 3: QO mediated overexpression of TUSC2 A,B, significantly decreases cell

viability (120 hours) in ALK+ cell lines that are resistant to Alectinib. All data have been analyzed using t-test. ****:p<0.0001,*:0.01≤p≤ 0.05.

✱✱✱✱

600000

400000

200000

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80000

60000

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20000

ALK192-P-03

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ALK192-P-01

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QO mediated TUSC2 overexpression can reduce cell

viability in ALK+ cells resistant to ALK inhibitor, Alectinib

CUTO 9

✱✱✱✱

CUTO 8

NCI-H2228

QO can overexpress TUSC2 in EML4-ALK+ NSCLC cell

lines and patient derived organoids (PDOs)

QO +

Alectinib

QO +

DMSO

Blank

Nanoparticles (BNP) + Alectinib

Blank

Nanoparticles (BNP) + DMSO

In-vitro Study design

QO in combination with ALK inhibitor, Alectinib, can induce

cell death

Cmax

6000

✱✱✱✱

4000

NCI-H2228

Start low

Start IC50

2000 ✱

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CUTO 8

CUTO 29.1

150

caspase 3/7 activity

BNP

TUSC2

✱

24 48 72 96 120 144

hours

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BNP QO

TUSC2

150

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NCI-H2228

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✱✱✱

- - + - - + - - + - - +

TUSC2

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✱✱

400 ✱✱✱✱

✱

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parental Cmax Start low

Start IC50

NCI-H2228

Cmax

Start low Start IC50

✱✱

✱

300 ✱✱✱✱ ✱✱✱✱

QO - + - - + - - + - - + -

✱✱✱✱

BNP

+ - - + - - + - - + - -

100

NCI-H2228

200

✱✱✱✱

100

Cleaved caspase 3 Tubulin

TUSC2

Tubulin

Cmax Start low Start IC50

✱✱✱✱

QO in combination with Alectinib has contrasting effects on

Alectinib-sensitive and Alectinib-resistant models

In-vivo Study design & Timeline

Treatment

~ 3 weeks 4 weeks

Cell Tumors Injection randomized into

4 groups

Alectinib-sensitive model: NCI-H2228-derived xenograft

100

BNP+vehicle control

QO+vehicle control BNP+Alectinib ✱

QO+Alectinib

BNP+vehicle control

QO+vehicle control

BNP+Alectinib

QO+Alectinib

0 20 40 60 80 100

Clinical Relevance (%) = Best Outcome (QO+Alectinib) − Worst Outcome (BNP+Alectinib) × 100

Best Outcome (QO+Alectinib)

Alectinib-resistant model: ALK167 patient derived xenograft

BNP+vehicle control

QO+vehicle control

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BNP+vehicle control BNP+Alectinib

QO+vehicle control

QO+Alectinib

Combination of QO with Alectinib exhibit significant synergy.

Fig 5: A, B, D, Mice were treated for a duration of 30 days, and tumor measurements were recorded daily. C,E, Post-treatment, mice were monitored during a drug free period to determine their survival. Data analyzed using t-test.

***:0.0001≤p≤0.001, **:0.001≤p≤0.01, *:0.01≤p≤ 0.05.

The upregulation of TUSC2 by QO induces apoptosis in ALK+ NSCLC cells, including those resistant to Alectinib.

Combining QO with Alectinib further increases apoptosis and improves treatment outcomes, but the precise mechanism through which TUSC2 regulates cell signaling requires additional research.

This work was supported by the Judith Tam ALK Lung Cancer Initiative.

We thank Genprex, Inc. for providing the Quaratusugene Ozeplasmid (QO) and the TUSC2 plasmid, and UC Anschutz for providing the CUTO cell lines.

We thank ALK Positive and Genprex, Inc. for support with in-vivo studies.

Travel to San Diego, CA for AACR 2026 was supported in-part by Rogel Cancer Center Postdoctoral Travel Award.

Group 4

QO + Alectinib

Group 3

BNP + Alectinib

Group 2

QO + DMSO

Group 1

BNP + DMSO

Alectinib-resistant model

Alectinib-sensitive model

Alectinib & vehicle control :

0.5mg/kg Daily x 30 days, oral gavage (sensitive model) 15mg/kg Daily x 30 days, oral gavage (resistant model).

QO & BNP :

25g/mouse, every 3rd day x 30 days, tail-vein injection

Survival study

~ 8 weeks

QO with Alectinib has a 23.5% improved outcome than Alectinib alone.

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Relative mRNA expression of TUSC2

Relative Cell Viability (%)

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

Treatment

Tumor shrinkage

QO + vehicle control

78.8%

BNP + Alectinib

60.14%

QO + Alectinib

78.60%

Background: Non-Small Cell Lung Carcinoma (NSCLC) with the EML4-ALK fusion gene represents about 5% of all NSCLC cases. While these tumors initially respond to ALK Tyrosine Kinase Inhibitors (TKIs), which are standard first- and second-line treatments, resistance develops in nearly all patients, creating an urgent need for new therapeutic strategies. This study investigates the potential of quaratusugene ozeplasmid (QO), a novel gene therapy developed by Genprex that encapsulates the Tumor Suppressor Candidate 2 (TUSC2) gene in non-viral lipid nanoparticles. TUSC2 is typically expressed at low levels in NSCLC, and its upregulation by QO was evaluated in a variety of ALK+ cell lines and patient-derived organoids (PDOs) before and after QO treatment.

Luminiscence

Relative % of colony formation

Relative Cell Viability (%)

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

Control

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

Methods and Results: The results demonstrated that QO-induced TUSC2 overexpression triggered strong pro-apoptotic responses in both ALK+ cells sensitive to and resistant (lab-generated) to the ALK inhibitor alectinib, as shown by increased apoptotic markers and decreased cell viability, especially when QO was combined with alectinib. To further explore this combination, two in vivo mouse models were used: one with alectinib-sensitive NCI-H2228 ALK+ cells and another with alectinib-resistant ALK167 PDX implants. Mice were divided into four groups: vehicle control, QO alone, alectinib alone (0.5 mg/kg for sensitive or 15 mg/kg for resistant, oral, daily), and QO plus alectinib. In the sensitive model, alectinib alone caused 60% tumor shrinkage, but QO alone and in combination with alectinib achieved 79% tumor reduction (p = 0.0135 vs. control), showing a 23% improvement over alectinib alone.

Relative % of BRDU+ cells

Major new unpublished results: In the resistant model, the QO

✱✱✱✱

Cmax

parental

Relative % of colony formation

Luminiscence

Relative Cell Viability (%)

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

and alectinib combination showed a synergistic effect, leading to the greatest tumor reduction and improved overall survival (p = 0.0001 vs. control), underscoring the clinical promise of this approach. Collectively, these in vitro and in vivo findings support the advancement of QO-mediated TUSC2 upregulation as a strategy to limit tumor growth and overcome drug resistance in ALK+ NSCLC, warranting further development toward clinical trials.

Luminiscence

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

✱✱✱✱

Relative mRNA expression of TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

BNP

TUSC2

Cell lines: EML4-ALK+ NSCLC cell lines, NCI-H2228 parental & its corresponding Alectinib resistant cell lines generated in the lab, Cmax, Start low, Start IC50, CUTO 8, 9, 29.1.

✱✱✱✱

Luminiscence

Patient-derived

Luminiscence

material (PDM): Organoids derived from human tissue and plural effusion.

/BNP

Control

TUSC2

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

BNP+DMSO

BNP+Alectinib

QO+DMSO

QO+Alectinib

*This figure was made using BioRender.com.

Disclaimer

Genprex Inc. published this content on April 19, 2026, and is solely responsible for the information contained herein. Distributed via Public Technologies (PUBT), unedited and unaltered, on April 19, 2026 at 22:19 UTC.