Dual targeting of mitochondrial Lon peptidase 1 and chymotrypsin-like protease by small molecule BT317, as potential therapeutics in malignant astrocytoma
Abstract Malignant astrocytomas are aggressive glioma tumors characterized by extensive hypoxia-induced, mitochondria-dependent changes such as altered respiration, increased chymotrypsin-like (CT-L) proteasome activity, decreased apoptosis, drug resistance, stemness and increased invasiveness. Mitochondrial Lon Peptidase I (LonP1) overexpression and increased CT-L proteasome inhibitors activity are the biomarkers of aggressive high grade glioma phenotype, poor prognosis and found to be associated with recurrence and poor patient survival, and drugs targeting either LonP1 or the CT-L activity have anti-glioma activity in preclinical models. We here for the first time introduced and evaluated a novel small molecule, BT317, derived from coumarinic compound 4 (CC4) using structure-activity modeling which we found to inhibit both LonP1 and CT-L proteasome activity. Using gain-of-function and loss-of-function genetic models, we discovered that BT317 is more effective than the individual LonP1 or CT-L inhibition in increasing reactive oxygen species (ROS) generation and inducing apoptosis in high-grade astrocytoma lines.In vitro, BT317 had activity as a single agent but, more importantly, enhanced synergy with the standard of care commonly used chemotherapeutic temozolomide (TMZ). In orthotopic xenograft, patient derived glioma models, BT317 was able to cross the blood-brain barrier, to show selective activity at the tumor site and to demonstrate therapeutic efficacy both as a single agent and in combination with TMZ. BT317 defines an emerging class of dual LonP1, and CT-L proteasome inhibitors exhibited promising anti-tumor activity and could be a promising candidate for clinical translation in the space of malignant astrocytoma therapeutics.Simple Summary We demonstrate here that BT317 has activity in patients derived malignant astrocytomas models by dual inhibiting mitochondrial Lon Peptidase I (LonP1) and the Chymotrypsin-Like (CT-L) proteasome activity. Malignant astrocytomas (including both IDH mutant astrocytomas grade 4 and IDH wildtype glioblastoma) have poor clinical outcomes, and novel treatments are needed to limit recurrence and improve overall survival. These tumors have a malignant phenotype that is mediated by altered mitochondrial metabolism, abnormal protein processing, and adaptation to hypoxia. We have previously published that gliomas are especially vulnerable to proteasome inhibitors as well as to inhibitors of the mitochondrial Lon Peptidase I (LonP1), but the effect of combining the two strategies has not been reported. Here, we present evidence dual inhibition of LonP1 and Chymotrypsin-like (CT-L) proteasome activity with the small-molecule BT317 can effectively induce cellular reactive oxygen species (ROS) production leading to apoptosis in clinically relevant malignant astrocytoma patient-derived orthotopic models. BT317 showed strong synergy with the standard of care, temozolomide (TMZ), in the astrocytoma models independent of their IDH profile. This preclinical study demonstrated a potential dual LonP1 and CT-L proteasome inhibitor as novel therapeutic strategies for malignant astrocytoma and provided insight for future clinical translation studies alone or in combination with the standard of care.Graphical Abstract <jats:fig id="ufig1" position="float" fig-type="figure" orientation="portrait"><jats:graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="536816v2_ufig1" position="float" orientation="portrait" /></jats:fig>Highlights <jats:list list-type="order">The novel compound BT317 acts as a dual inhibitor of LonP1 and chymotrypsin-like proteasome enzymatic activity.LonP1 and CT-L proteasome inhibition by BT317 drives ROS production in malignant astrocytoma independent of genetic profile.LonP1 and CT-L proteasome inhibition drives autophagy in IDH mutant astrocytoma.BT317 shows blood-brain barrier permeability and has low normal tissue toxicity.BT317 synergizes with the first-line chemotherapy agent Temozolomide (TMZ)..
| Medienart: |
|
|---|
Preprint| Erscheinungsjahr: |
2024 |
|---|---|
| Erschienen: |
2024 |
| Enthalten in: |
bioRxiv.org - (2024) vom: 16. Apr. Zur Gesamtaufnahme - year:2024 |
|---|
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Douglas, Christopher [VerfasserIn] |
|---|
| Links: |
Volltext [kostenfrei] |
|---|
| Themen: |
|---|
| doi: |
10.1101/2023.04.13.536816 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
XBI039280853 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | XBI039280853 | ||
| 003 | DE-627 | ||
| 005 | 20240419090806.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 230418s2024 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1101/2023.04.13.536816 |2 doi | |
| 035 | |a (DE-627)XBI039280853 | ||
| 035 | |a (biorXiv)10.1101/2023.04.13.536816 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Douglas, Christopher |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Dual targeting of mitochondrial Lon peptidase 1 and chymotrypsin-like protease by small molecule BT317, as potential therapeutics in malignant astrocytoma |
| 264 | 1 | |c 2024 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 520 | |a Abstract Malignant astrocytomas are aggressive glioma tumors characterized by extensive hypoxia-induced, mitochondria-dependent changes such as altered respiration, increased chymotrypsin-like (CT-L) proteasome activity, decreased apoptosis, drug resistance, stemness and increased invasiveness. Mitochondrial Lon Peptidase I (LonP1) overexpression and increased CT-L proteasome inhibitors activity are the biomarkers of aggressive high grade glioma phenotype, poor prognosis and found to be associated with recurrence and poor patient survival, and drugs targeting either LonP1 or the CT-L activity have anti-glioma activity in preclinical models. We here for the first time introduced and evaluated a novel small molecule, BT317, derived from coumarinic compound 4 (CC4) using structure-activity modeling which we found to inhibit both LonP1 and CT-L proteasome activity. Using gain-of-function and loss-of-function genetic models, we discovered that BT317 is more effective than the individual LonP1 or CT-L inhibition in increasing reactive oxygen species (ROS) generation and inducing apoptosis in high-grade astrocytoma lines.In vitro, BT317 had activity as a single agent but, more importantly, enhanced synergy with the standard of care commonly used chemotherapeutic temozolomide (TMZ). In orthotopic xenograft, patient derived glioma models, BT317 was able to cross the blood-brain barrier, to show selective activity at the tumor site and to demonstrate therapeutic efficacy both as a single agent and in combination with TMZ. BT317 defines an emerging class of dual LonP1, and CT-L proteasome inhibitors exhibited promising anti-tumor activity and could be a promising candidate for clinical translation in the space of malignant astrocytoma therapeutics.Simple Summary We demonstrate here that BT317 has activity in patients derived malignant astrocytomas models by dual inhibiting mitochondrial Lon Peptidase I (LonP1) and the Chymotrypsin-Like (CT-L) proteasome activity. Malignant astrocytomas (including both IDH mutant astrocytomas grade 4 and IDH wildtype glioblastoma) have poor clinical outcomes, and novel treatments are needed to limit recurrence and improve overall survival. These tumors have a malignant phenotype that is mediated by altered mitochondrial metabolism, abnormal protein processing, and adaptation to hypoxia. We have previously published that gliomas are especially vulnerable to proteasome inhibitors as well as to inhibitors of the mitochondrial Lon Peptidase I (LonP1), but the effect of combining the two strategies has not been reported. Here, we present evidence dual inhibition of LonP1 and Chymotrypsin-like (CT-L) proteasome activity with the small-molecule BT317 can effectively induce cellular reactive oxygen species (ROS) production leading to apoptosis in clinically relevant malignant astrocytoma patient-derived orthotopic models. BT317 showed strong synergy with the standard of care, temozolomide (TMZ), in the astrocytoma models independent of their IDH profile. This preclinical study demonstrated a potential dual LonP1 and CT-L proteasome inhibitor as novel therapeutic strategies for malignant astrocytoma and provided insight for future clinical translation studies alone or in combination with the standard of care.Graphical Abstract <jats:fig id="ufig1" position="float" fig-type="figure" orientation="portrait"><jats:graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="536816v2_ufig1" position="float" orientation="portrait" /></jats:fig>Highlights <jats:list list-type="order">The novel compound BT317 acts as a dual inhibitor of LonP1 and chymotrypsin-like proteasome enzymatic activity.LonP1 and CT-L proteasome inhibition by BT317 drives ROS production in malignant astrocytoma independent of genetic profile.LonP1 and CT-L proteasome inhibition drives autophagy in IDH mutant astrocytoma.BT317 shows blood-brain barrier permeability and has low normal tissue toxicity.BT317 synergizes with the first-line chemotherapy agent Temozolomide (TMZ). | ||
| 650 | 4 | |a Biology |7 (dpeaa)DE-84 | |
| 650 | 4 | |a 570 |7 (dpeaa)DE-84 | |
| 700 | 1 | |a Jain, Shashi |e verfasserin |0 (orcid)0000-0001-9428-0135 |4 aut | |
| 700 | 1 | |a Lomeli, Naomi |e verfasserin |0 (orcid)0000-0002-1071-386X |4 aut | |
| 700 | 1 | |a Di, Kaijun |e verfasserin |4 aut | |
| 700 | 1 | |a Nandwana, Nitesh Kumar |e verfasserin |4 aut | |
| 700 | 1 | |a Mohammed, Adil Shareef |e verfasserin |4 aut | |
| 700 | 1 | |a Vu, Thao |e verfasserin |4 aut | |
| 700 | 1 | |a Pham, James |e verfasserin |4 aut | |
| 700 | 1 | |a Lepe, Javier |e verfasserin |4 aut | |
| 700 | 1 | |a Kenney, Maria Cristina |e verfasserin |4 aut | |
| 700 | 1 | |a Das, Bhaskar |e verfasserin |4 aut | |
| 700 | 1 | |a Bota, Daniela A. |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t bioRxiv.org |g (2024) vom: 16. Apr. |
| 773 | 1 | 8 | |g year:2024 |g day:16 |g month:04 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1101/2023.04.13.536816 |x 0 |z kostenfrei |3 Volltext |
| 912 | |a GBV_XBI | ||
| 951 | |a AR | ||
| 952 | |j 2024 |b 16 |c 04 | ||