For its next generation T-cell product candidates, the company will incorporate multiple modular enhancements, but as the authors of this point out engineering multiple features into one cell product is limited by the constraints of current viral vectors, which leads to a heterogeneous population of cells.
Using a leucine zipper-based cell sorting enabled a selective single-step purification of cells co-transduced with two vectors, designed to potentially double the number of incorporated transgenes. This facilitated generation of T-cells simultaneously expressing up to four CARs (CD19, CD20, CD79b, and BAFF-R) and co-expressing up to three switch receptors (CD200R-CD27, PD-1-OX40, and Fas-41BB) https://www.biorxiv.org/content/10.1101/2023.09.13.557232v1
AUTO4 has a unique targeting mechanism. Mature T-cells express either T-cell receptor B-chain constant domains 1 or 2 (TRBC1 or TRBC2). T-cell lymphomas are clonal and either express TRBC1 or TRBC2. So a targeting strategy based on mutually exclusive expression of TRBC domains can spare a proportion of the normal T-cell compartment.
CART is like a big bomb while carpet bombing with bomblets is needed to rid cancer everywhere in the body.
Rare and aggressive group of NHLs without the benefit of B-cell targeted treatments. Few options after CHOP like regimens
A privilege to be involved in this first in human #CAR-T trial in R/R #PTCL - an area of unmet need.— Kate Cwynarski (@CwynKate) June 15, 2023
Promising Ph I safety data- no ICANS, only 1/13 Gr3 CRS
& encouraging efficacy at highest dose: CMR in 2/4 patients w/ highest dose at 12 & 15 months#17ICML @gloria_iacoboni https://t.co/edUCXhI1tx
Another oral presentation is planned for a different (peer-reviewed) medical/scientific even, with long-term follow-up planned for ASH. The company is targeting the BLA submission for the program towards the end of this year, MAA filing towards the end of the first quarter of 2024, and the UK filing in the second quarter of next year.
Safety and efficacy of obecabtagene autoleucel (obe-cel, AUTO1), a fast-off rate CD19 CAR, in relapsed/refractory adult B-cell acute lymphoblastic leukemia (r/r B-ALL): Top line results of the pivotal FELIX study https://meetings.asco.org/abstracts-presentations/219864
3318 Safety, Efficiency and Long-Term Follow-up of AUTO1, a Fast-Off Rate CD19 CAR in Relapsed/Refractory B-Cell Acute Lymphoblastic Leukaemia and Other B-Cell Malignancies https://ash.confex.com/ash/2022/webprogram/Paper167053.html
4650 Dual Antigen Targeting with Co-Transduced CD19/22 CAR T Cells May Prevent Antigen-Negative Relapse after CAR T Cell Therapy for Relapsed/Refractory ALL https://ash.confex.com/ash/2022/webprogram/Paper164879.html
4634 First in Human Study of AUTO4, a TRBC1-Targeting CAR T-Cell Therapy in Relapsed/Refractory TRBC1-Positive Peripheral T-Cell Lymphoma https://ash.confex.com/ash/2022/webprogram/Paper165971.html
258 A Novel Protein-Based Approach to Generate Allogeneic CAR-T Cells with Simultaneous TCR and MHC Class 1 Downregulation https://ash.confex.com/ash/2022/webprogram/Paper167980.html
I found out that a Chinese group  is doing a similar allo approach to the company's , with some promising clinical data. So far, no GvHD and a high response rate.
I found this
AUTL presented three posters at ASGCT over the last two days. Most relevant near term to AUTL is the CCR poster, as AUTO6NG (GD2), which is scheduled to enter the clinic this year, contains an IL-7-CCR. The poster noted that gene expression from an IL-7-CCR-GD2CAR-T was largely identical to both an IL-2 and IL-12-CCR-GD2CAR-T. Notably in mice a GM-CSF-CCR-GD2CAR-T produced the greatest CAR-T expansion, persistence and overall survival.
In addition to a lack of cytokine support in the solid tumour microenvironment, FasL expression could trigger killing of activated CAR-T since these cells will express Fas. AUTL described a Fas-CD40 construct that neutralised FasL induced apoptosis while maintaining CAR-T function. Controlled activation of CAR and or cytokine secretion may be advantageous in solid tumours and AUTL describes a two component system based on minocycline. For a CAR, minocycline inactivates the CAR while for IL-12, use of minocycline leads to secretion.
Cytokine support of cell therapy has most often required systemic dosing of cytokines. A chimeric chemokine receptor with constant heavy and light chain domains from an IgG are fused to the transmembrane and intracellular domains of the chosen cytokine receptor. An IL-2 CCR drove significant T-cell expansion in the absence of cytokine starvation CCR control
Of the top fifty genes activated by exogenous IL-2, forty-four were activated by IL-2-CCR. Truncation of the IL2R beta chain resulted in either increased or diminished T cell expansion. Substitution of IL-7 for IL-2, increased T-cell expansion, while IL-18 increased IFNy secretion over IL-2. Substitution of GM-CSF for IL-2 increased myeloid cell expansion.
Eighteen CCR-GD2 CARs were evaluated against a GD2 CAR control and while most preserved CAR-T effector function, the IL-12 and IL-7 CCR's drove greater CAR-T expansion and production of IFNy. Gene expression changes for IL-2-CCR and IL-7-CCR and IL-7-CCR and IL-12-CCR were identical.
In a GD-2 colon cancer model using murinised components, CCR-IL-7 or CCR-GM-CSF GD2 CAR-T, demonstrated improved tumour control compared to GD2 CAR-T; CAR-T expansion was significantly higher for the GM-CSF-CCR and correlated with a pronounced increase in overall survival.
In a metastatic melanoma model similar observations were made and at day fourteen, only GM-CSF-CCR-GD2 CAR-T were detected.
FAS: Fas ligand is expressed by most cancers, regulatory T-cells, endothelial cells, myeloid-derived suppressor cells and cancer-associated fibroblasts. Activated T-cells including CAR-T constitutively express Fas and engagement of FasL leads to CAR-T death via apoptosis. Truncated non signalling Fas rescued T-cells from Fas mediated apoptosis, but significantly diminished CAR-T proliferation and efficacy
Adding the intracellular pro-survival domain of a TNF receptor rescued CAR-T proliferation in the presence of FasL; seventeen different receptors were evaluated including a short of BCMA, 41BB, CD27, CD40 and Fn14.
Against GD2 FasL+ target cells, GD2 CAR-T exhibited a high degree of apoptosis which was reversed in the presence of Fas truncated or Fas-receptors. While all GD2 CAR-T with one of five shortlisted Fas combinations were equipped against GD2+ target cells, with repeat stimulation, Fas-CD40 retained the highest level of activity, correlating with greater retention of a central memory phenotype.
1105: CAR-T Cells Engineered to Express a FAS-CD40 Chimera Display Superior Persistence and Tumour Cytotoxicity https://annualmeeting.asgct.org/abstracts/abstract-details?abstractId=2004
1096: Enhancing CAR T Cell Therapy Using Fab Based Constitutively Heterodimeric Cytokine Receptors https://annualmeeting.asgct.org/abstracts/abstract-details?abstractId=1996
311: Development of a Minocycline Mediated Protein-Protein Displacement Platform Using an Anti-Minocycline Single Domain Antibody and a Dedicated Displaceable Peptide https://annualmeeting.asgct.org/abstracts/abstract-details?abstractId=1267
CAR-T Cells Engineered to Express a FAS-CD40 Chimera Display Superior Persistence and Tumour Cytotoxicity.
Enhancing CAR T Cell Therapy Using Fab Based Constitutively Heterodimeric Cytokine Receptors.
Development of a Minocycline Mediated Protein-Protein Displacement Platform Using an Anti-Minocycline Single Domain Antibody and a Dedicated Displaceable Peptide.
Also, clinical data from AUTO4*.
* Trial enrolment has been upsized from 55 to 200, completion delayed from Q2 2022 to Q3 2023.
A High Sensitivity aCD22 CAR Combined with aCD19 CAR to Generate Dual Targeting CAR T Cells for the Treatment of r/r B-ALL https://ash.confex.com/ash/2021/webprogram/Paper152655.html
Industrialization of an Academic Miltenyi Prodigy-Based CAR T Process https://ash.confex.com/ash/2021/webprogram/Paper153031.html
Safety and Efficacy of AUTO1, a Fast-Off Rate CD19 CAR in Relapsed/Refractory B-Cell Non-Hodgkin's Lymphoma (B-NHL) and Chronic Lymphocytic Leukaemia (CLL) https://ash.confex.com/ash/2021/webprogram/Paper146316.html
Two ways to potentially improve the activity of AUTO6NG in either neuroblastoma or osteosarcoma https://onlinelibrary.wiley.com/doi/10.1111/cas.15074 https://cancerimmunolres.aacrjournals.org/content/4/10/869.long
The company announced an agreement with MRNA, granting them an exclusive license to develop and commercialise mRNA therapeutics incorporating proprietary binders for up to four IO targets. AUTL would be eligible to receive an upfront payment for each target licensed and development and commercial milestone payments for each product successfully commercialised. In addition, entitled to receive royalties on net sales of all products commercialised under the agreement.
Dr. Itin, will participate in a panel discussion titled ''Cell Therapies In the Next Decade'' to be held on Wednesday, July 14, at 08.55 a.m. EDT at the William Blair Biotech Focus Conference, taking place virtually from July 14-15. The company will also host virtual one-on-one meetings at the conference. The live panel discussion can be viewed from the investor section of the website and will be available for a period of 30 days after the conference.
As of the data cut-off date of May 17, thirteen patients in Cohort D with R/R IBCL had been enrolled in the trial and product was successfully manufactured for twelve patients, with one patient’s cells ongoing in manufacture. As of the data cut-off date, nine had received AUTO1 infusion. Three were pending infusion (including the patient above) and one patient died prior to lymphodepletion due to a Cov infection. Obe-cel was well tolerated and demonstrated a favourable safety profile, despite high disease burden. All treated patients achieved a complete metabolic response and had robust CAR-T engraftment, expansion, and persistence.
Grade 1 CRS was reported in four patients and Grade 2 CRS in one. No immune effector cell-associated neurotoxicity syndrome of any grade was observed. At a median follow-up of six months (range 4.0-8.1), eight of nine patients were disease free, with one relapse at month six, but was rescued with radiotherapy. One died of a Cov infection at month 5.6 whilst in complete metabolic response.
Also, twenty patients in Cohort A with R/R aALL had received obe-cel. The therapy was well tolerated, with no patients experiencing Grade 3 or higher CRS. Three patients, all of whom had high leukaemia burden (over 50% blasts), experienced Grade 3 ICANS that resolved swiftly with steroids.
Of the twenty patients evaluable for efficacy, seventeen achieved minimum residual disease negative complete response at one month. Most notably, the durability of remissions is highly encouraging. Across all treated patients, event free survival at twelve months and twenty-four months is 50.2% with the median not being reached.
The call https://edge.media-server.com/mmc/p/wbmik6fg
AUTO1 CAR-T cells from pediatric ALL patients who still had CAR-T cells detectable in the blood more than two years after their treatment were compared with patients who had lost their AUTO1 CAR-T cells one to two months post treatment. The study shows that a subset of AUTO1 CAR-T cells called stem cell memory T-cells appear critical in both the initial anti-leukemic response and for long term immune surveillance. This suggests that this sub-group of AUTO1 CAR-T cells contribute to the long-term durability of effect that AUTO1 has in these patients.
''AUTO1 has been designed to have an optimized interaction between its chimeric antigen receptor and the CD19 target on cancer cells,'' said Dr Martin Pule (Founder and CSO). ''This means AUTO1 can efficiently deliver a kill and disengage rapidly like a normal T-cell, leading to less exhaustion and less T-cell differentiation. This unique property of AUTO1 potentially contributes to the enrichment and maintenance of this stem cell memory subset that appears to be critical to the long-term durability observed in pediatric ALL patients treated with AUTO1.''
The paper https://www.nature.com/articles/s43018-021-00207-7
So far, so good. If this holds up, it could be best-in-class https://www.globenewswire.com/news-release/2021/05/12/2228342/0/en/Autolus-Therapeutics-to-Present-New-Data-on-AUTO1-in-r-r-Indolent-B-Cell-Lymphomas-at-the-European-Hematology-Association-Virtual-Congress.html
Wells Fargo Biotech Cell and Gene Therapy Companion o Newsletter. This week's themes include: continued fund-raising for cell and gene therapy; regulatory actions including first BCMA CAR-T product approval, a first CAR-T filing for adult acute lymphoblastic leukemia, IND approval for pre-loaded NK cells, Orphan Drug designation in TTR-amyloidosis; optimization of cell therapy including point-of-care, reversal of exhaustion; further exoneration of AVV as a driver of cancer, but caution over long-term toxicity from AAV9-SMN in mice with spinal muscular atrophy; funding of an academic CRISPR trial in sickle cell disease and pre-clinical data for a gene therapy addressing hemophilia A & B.
Highlights of the week include: 1) Applied Genetic Technologies Corporation (AGTC) partner, Bionic Sight disclosing positive data for BS01 in retinitis pigmentosa. 2) Emergence of potential point-of-care cell therapy competitor, Avalon GloboCare - relevant for Precigen (PEGN) and Ziopharm (ZIOP). 3) Gracell Biotechnologies (GRCL) agreement with Lonza to manufacture dual CD19/BCMA FasCAR-T, GC-012F.4) Announcement by uniQure (QURE) that independent investigation finds it highly unlikely that the case of hepatocellular carcinoma was caused by AAV. 5) Publication with senior author Guangping Gao that natural AAV sequences in tumors and adjacent tissue are not different from each other - relevant for QURE and Sangamo Therapeutics (SGMO). 6) IND approval for Affimed's (AFMD) first IND for pre-loaded NK cell therapy. 7) Autolus Therapeutic's (AUTL) obe-cel received PRIME designation for relapsed/refractory acute lymphoblastic leukemia (ALL). (8) Filing of an sBLA by Gilead for TECARTUS in adult ALL highlights obe-cel's superior safety profile. 9) FDA approved bluebird bio (BLUE) and BMY's BCMA CAR-T, ABECMA in a more restricted patient population than studied - relevant for AUTL, Cellectis (CLLS), Celyad Oncology (CYAD), Fate Therapeutics (FATE), GRCL. 9) Intellia Therapeutics (NTLA) received Orphan Drug designation in Europe for NTLA-2001 in TTR-amyloidosis. 10) CIRM will fund a CRISPR gene therapy trial in sickle cell disease at 3 University of California institutions - relevant for Beam Therapeutics (BEAM), BLUE, CRISPR Therapeutics (CRSP) & SGMO. 11) UCLA publishes pre-clinical data supporting clinical development of CD4 CAR-T for HIV; NIH publishes less encouraging data [this is science]. 12) Clinical vignette from Chinese glioblastoma patient treated with a B7H3 CAR-T published - relevant for FATE. 13) Columbia University publish pre-clinical data suggesting gain of toxic function data in mice receiving AAV9 SMN treatment for spinal muscular atrophy - relevant for Ionis Pharmaceuticals (IONS) if there is read through to NVS's ZOLGENSMA. 14) Pre-clinical gene therapy with a platelet restricted factor Xa is effective in hemophilia A & B - relevant to QURE/SGMO. 15) CAR-T exhaustion can be reversed in pre-clinical models with a single dose of dasatinib - relevant for all companies developing CAR-T using a high affinity/avidity scFv.
Notable Corporate Events
AGTC partner Bionic Sight reported initial data from a phase 1/2 study of BS01 in advanced stage retinitis pigmentosa (RP). The data demonstrated that all four patients, who were complete or near-completely blind, can now see light and motion. Additionally, 2/4 patients can now detect the direction of motion. Bionic expects to report additional data later in 2021. Recall, AGTC has the option to pick up the program when the phase 1/2 data are available.
Avalon GloboCare Corp and strategic partner Arbele, are collaborating with the University of Pittsburgh Medical Center (UPMC) Hillman Cancer Center to advance the clinical development of point of care RNA-based autologous cell therapies using FLASH CAR technology. Avalon’s first FLASH CAR product candidate, AVA-011 targets CD19 and CD22 is on track to initiate a clinical study in B-cell leukemia and non-Hodgkin’s lymphoma – relevant for PEGN and ZIOP who use on-site point of care CAR-T manufacture leveraging Sleeping Beauty. PEGN and ZIOP use conventional DNA-CARs which integrate into the T cell genome while Avalon is using an RNA CAR which may be episomal and shorter lived.
GRCL signed an agreement with Lonza to manufacture FasTCAR products to support IND filing of dual BCMA/CD19 FasTCAR, GC-012F. Separately, GRCL announced enrollment of the first acute lymphoblastic leukemia patient into the registration-directed trial of GC007g in China. GC007g is an HLA-matched donor-derived allogeneic CD19 CAR-T product.
QURE announced the results of an independent investigation into the case of hepatocellular carcinoma (HCC) diagnosed in a patient in the HOPE-B trial. QURE noted that the independent investigation found it highly unlikely the HCC was caused by the company's AAV gene therapy etranacogene dezaparvovec (etra-dez).
Notable Regulatory/Clinical News
AFMD and partner NKMax America announced FDA IND clearance for a co-sponsored phase 1/2a dose escalation and expansion study evaluating AFMD’s EGFR/CD16A innate cell engager AFM24 in combination with NKMax America’s autologous NK-cell product SNK-01, currently in phase 1 testing, in patients with advanced/metastatic EGFR-expressing solid tumors.
AUTL announced receipt of PRIME designation for Obe-cel (AUTO1) in relapsed/refractory B-acute lymphoblastic leukemia (B-ALL) as well as recent listing of the MCARTY trial (NCT04795882) evaluating a modular CAR-T targeting BCMA alone or BCMA with CD19.
AUTL – Filing of an sBLA by GILD for TECARTUS highlights the superior safety profile of obe-cel (AUTO-1). With 1/4 and nearly 1/2 of TECARTUS-treated adult acute lymphoblastic leukemia patients suffering grade 3 or higher cytokine release syndrome (CRS) and neurotoxicity / immune cell associated neurotoxicity syndrome (ICANS) respectively at the 1M cell dose of ZUMA-3. By contrast, in 20 patients treated with obe-cel, AUTL reported no grade 3 or higher CRS and ICANS was limited to 4/20 patients, of which 3 cases were grade 1 and resolved within 24 hours.
AUTL – Chinese academics listed a phase 1 trial of a TRBC1 CAR-T in TRBC1+ve relapsed/refractory T cell leukemia. AUTL is conducting the LIbrA T1 trial of AUTO4 in TRBC1+ve T cell leukemia with interim data expected 2H21; AUTL expects to initiate clinical development of AUTO5 in TRBC2 T cell leukemia 2H21.
AUTL/CLLS/CYAD/ FATE & GRCL - we see the approved label for bluebird bio/ Bristol Myers Squibb's BCMA CAR-T, ABECMA for relapsed/refractory multiple myeloma setting a low bar for competitors. While ABECMA was studied in 4th line plus myeloma FDA gave the product a 5th line plus label. ABECMA’s blackbox warning includes hemophagocytic lymphohistiocytosis/macrophage activation syndrome (HLH/MAS) which occurred in 4% of patients overall increasing to 8% at the 450MM dose; approved CD19 CAR-T do not have an HLH/MAS blackbox warnings. In more recent BCMA trials, we note use of the IL-1 receptor antagonist, ANKINRA, potentially for HLH/MAS with 19% use in JNJ/Legend Biotech's cilta-cel CARTITUDE-I trial.
NTLA announced receipt of orphan drug designation from European Commission (EC) for Regeneron-partnered in vivo editing program NTLA-2001 for TTR amyloidosis (ATTR). The decision follows the initiation of the phase 1 study of NTLA-2001 in hereditary ATTR with polyneuropathy (hATTR-PN).
BEAM/BLUE/CRSP/EDIT - California Institute of Regenerative Medicine will support a phase 1 trial of CRISPR engineered SCD gene therapy to be conducted at The University of California, San Francisco (UCSF), UC Berkeley (UCB) and UC Los Angeles (UCLA).The 4-year study will include six adults and three adolescents with severe sickle cell disease. It is planned to begin this summer in Oakland and Los Angeles.
Notable Peer Reviewed Publications
FATE/CLLS – With both companies expected to announce programs outside of oncology we note UCLA scientists publish data for a 2nd generation anti-HIV1 CAR-T. UCLA’s use of engineered hematopoietic stem cells could be replaced by FATE’s induced pluripotent stem cells in our view. UCLA modified hematopoietic stem cells with a truncated CD4 CAR to target HIV envelope, that unlike the first gen CAR prevented HIV transmission as well as prevented IL-16 binding and potential non-specific CAR signaling while maintaining similar cytotoxicity against Env+ve cells. The use of 41BB as a costimulatory domain of the CAR, versus CD28 was essential for successful differentiation and improved anti-viral function. Suppression of viremia in untreated mice by the 2nd gen CAR was enhanced by combination treatment with ART which increased CAR-T persistence. The authors believe the 2nd gen CAR-T should be pursued as a clinical candidate. Separately we note an NIH manuscript describing the development of CAR-T against SIV. The studies failed to protect or gain control of viral spread. The authors identified optimal targets noting that while minimally expanded T cells were the most potent, generating large numbers was a challenge, one we note would not be an issue for an iPSC/allogeneic CAR-T. In addition, the authors observed that virus infected cells expressed Env protein for a relatively small fraction of the rapid viral lifecycle, and only towards the end when virus may be about to be released, an observation we interpret as requiring CAR-T persistence or the ability to re-dose.
They are now working on ways to further improve ex vivo expansion (using an inhibitor of the PI3K/AKT/mTOR pathway, platelet lysate and/or IL-21 [1-7]), as well as engineering some their CAR-T cell products to locally secrete anti-ectoenzyme antibodies to either CD39* and CD73*, and/or GSI**.
* High adenosine (ADO) in the tumour microenvironment suppresses the immune response by inhibiting antitumour immune functions (cytotoxic and effector functions of T- and NK cells) and promoting the development of immunosuppressive cells (Tregs, MDSCs, TAMs and/or tolerogenic DCs). Extracellular ADO can be generated from ATP released by cancer cells undergoing stress and/or death through the combined actions of CD39 (ATP to AMP) and CD73 (AMP to ADO).
** https://ashpublications.org/blood/article/134/19/1585/374996/Secretase-inhibition-increases-efficacy-of-BCMA https://ashpublications.org/blood/article/134/Supplement_1/1856/427564/Response-to-Bcma-CAR-T-Cells-Correlates-with